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2024
PJ Thiele, R Mela-Lopez, SA Blandin, D Klug
Let it glow: genetically encoded fluorescent reporters in Plasmodium Journal Article
In: Malaria Journal, vol. 23, no. 114, 2024.
Abstract | Links | BibTeX | Tags: blandin, fluorescent reporters, M3i, Plasmodium
@article{PJ2024,
title = {Let it glow: genetically encoded fluorescent reporters in Plasmodium},
author = {Thiele PJ and Mela-Lopez R and Blandin SA and Klug D},
editor = {Springer Nature},
doi = {10.1186/s12936-024-04936-9},
year = {2024},
date = {2024-04-20},
urldate = {2024-04-20},
journal = {Malaria Journal},
volume = {23},
number = {114},
abstract = {The use of fluorescent proteins (FPs) in Plasmodium parasites has been key to understand the biology of this obligate intracellular protozoon. FPs like the green fluorescent protein (GFP) enabled to explore protein localization, promoter activity as well as dynamic processes like protein export and endocytosis. Furthermore, FP biosensors have provided detailed information on physiological parameters at the subcellular level, and fluorescent reporter lines greatly extended the malariology toolbox. Still, in order to achieve optimal results, it is crucial to know exactly the properties of the FP of choice and the genetic scenario in which it will be used. This review highlights advantages and disadvantages of available landing sites and promoters that have been successfully applied for the ectopic expression of FPs in Plasmodium berghei and Plasmodium falciparum. Furthermore, the properties of newly developed FPs beyond DsRed and EGFP, in the visualization of cells and cellular structures as well as in the sensing of small molecules are discussed.},
keywords = {blandin, fluorescent reporters, M3i, Plasmodium},
pubstate = {published},
tppubtype = {article}
}
The use of fluorescent proteins (FPs) in Plasmodium parasites has been key to understand the biology of this obligate intracellular protozoon. FPs like the green fluorescent protein (GFP) enabled to explore protein localization, promoter activity as well as dynamic processes like protein export and endocytosis. Furthermore, FP biosensors have provided detailed information on physiological parameters at the subcellular level, and fluorescent reporter lines greatly extended the malariology toolbox. Still, in order to achieve optimal results, it is crucial to know exactly the properties of the FP of choice and the genetic scenario in which it will be used. This review highlights advantages and disadvantages of available landing sites and promoters that have been successfully applied for the ectopic expression of FPs in Plasmodium berghei and Plasmodium falciparum. Furthermore, the properties of newly developed FPs beyond DsRed and EGFP, in the visualization of cells and cellular structures as well as in the sensing of small molecules are discussed.
I Iacobucci, V Monaco, A Hovasse, B Dupouy, R Keumoe, B Cichocki, M Elhabiri, B Meunier, JM Strub, M Monti, S Cianférani, SA Blandin, C Schaeffer-Reiss, E Davioud-Charvet
Proteomic Profiling of Antimalarial Plasmodione Using 3-Benz(o)ylmenadione Affinity-Based Probes Journal Article
In: Chembiochem, 2024.
Abstract | Links | BibTeX | Tags: Activity-Based Protein Profiling, Antiprotozoal agents, Biological Chemistry, blandin, Chemical Biology, M3i, Photoaffinity Labeling, proteomics
@article{I2024,
title = {Proteomic Profiling of Antimalarial Plasmodione Using 3-Benz(o)ylmenadione Affinity-Based Probes},
author = {Iacobucci I and Monaco V and Hovasse A AND Dupouy B AND Keumoe R AND Cichocki B AND Elhabiri M AND Meunier B AND Strub JM AND Monti M AND Cianférani S AND Blandin SA AND Schaeffer-Reiss C AND Davioud-Charvet E},
editor = {Wiley-VCH GmbH},
url = {https://pubmed.ncbi.nlm.nih.gov/38639212/},
doi = {10.1002/cbic.202400187},
year = {2024},
date = {2024-04-19},
journal = {Chembiochem},
abstract = {Understanding the mechanisms of drug action in malarial parasites is crucial for the development of new drugs to combat infection and to counteract drug resistance. Proteomics is a widely used approach to study host-pathogen systems and to identify drug protein targets. Plasmodione is an antiplasmodial early-lead drug exerting potent activities against young asexual and sexual blood stages in vitro with low toxicity to host cells. To elucidate its molecular mechanisms, an affinity-based protein profiling (AfBPP) approach was applied to yeast and P. falciparum proteomes. New (pro-) AfBPP probes based on the 3-benz(o)yl-6-fluoro-menadione scaffold were synthesized. With optimized conditions of both photoaffinity labeling and click reaction steps, the AfBPP protocol was then applied to a yeast proteome, yielding 11 putative drug-protein targets. Among these, we found four proteins associated with oxidoreductase activities, the hypothesized type of targets for plasmodione and its metabolites, and other proteins associated with the mitochondria. In Plasmodium parasites, the MS analysis revealed 44 potential plasmodione targets that need to be validated in further studies. Finally, the localization of a 3-benzyl-6-fluoromenadione AfBPP probe was studied in the subcellular structures of the parasite at the trophozoite stage. },
keywords = {Activity-Based Protein Profiling, Antiprotozoal agents, Biological Chemistry, blandin, Chemical Biology, M3i, Photoaffinity Labeling, proteomics},
pubstate = {published},
tppubtype = {article}
}
Understanding the mechanisms of drug action in malarial parasites is crucial for the development of new drugs to combat infection and to counteract drug resistance. Proteomics is a widely used approach to study host-pathogen systems and to identify drug protein targets. Plasmodione is an antiplasmodial early-lead drug exerting potent activities against young asexual and sexual blood stages in vitro with low toxicity to host cells. To elucidate its molecular mechanisms, an affinity-based protein profiling (AfBPP) approach was applied to yeast and P. falciparum proteomes. New (pro-) AfBPP probes based on the 3-benz(o)yl-6-fluoro-menadione scaffold were synthesized. With optimized conditions of both photoaffinity labeling and click reaction steps, the AfBPP protocol was then applied to a yeast proteome, yielding 11 putative drug-protein targets. Among these, we found four proteins associated with oxidoreductase activities, the hypothesized type of targets for plasmodione and its metabolites, and other proteins associated with the mitochondria. In Plasmodium parasites, the MS analysis revealed 44 potential plasmodione targets that need to be validated in further studies. Finally, the localization of a 3-benzyl-6-fluoromenadione AfBPP probe was studied in the subcellular structures of the parasite at the trophozoite stage.
CF Estevez-Castro, MF Rodrigues, A Babarit, FV Ferreira, de Andrade EG, E Marois, R Cogni, ERGR Aguiar, JT Marques, RP Olmo
Neofunctionalization driven by positive selection led to the retention of the loqs2 gene encoding an Aedes specific dsRNA binding protein Journal Article
In: BMC Biol , vol. 22, no. 14, 2024.
Abstract | Links | BibTeX | Tags: Aedes mosquitoes, Double-stranded RNA (dsRNA), dsRNA binding protein (dsRBP), loqs2, M3i, marois, Marques, Olmo, RNA interference (RNAi)
@article{CF2024,
title = {Neofunctionalization driven by positive selection led to the retention of the loqs2 gene encoding an Aedes specific dsRNA binding protein},
author = {Estevez-Castro CF and Rodrigues MF and Babarit A and Ferreira FV and de Andrade EG and Marois E and Cogni R and Aguiar ERGR and Marques JT and Olmo RP},
url = {https://doi.org/10.1186/s12915-024-01821-4},
doi = {10.1186/s12915-024-01821-4},
year = {2024},
date = {2024-01-25},
urldate = {2024-01-25},
journal = {BMC Biol },
volume = {22},
number = {14},
abstract = {Background
Mosquito borne viruses, such as dengue, Zika, yellow fever and Chikungunya, cause millions of infections every year. These viruses are mostly transmitted by two urban-adapted mosquito species, Aedes aegypti and Aedes albopictus. Although mechanistic understanding remains largely unknown, Aedes mosquitoes may have unique adaptations that lower the impact of viral infection. Recently, we reported the identification of an Aedes specific double-stranded RNA binding protein (dsRBP), named Loqs2, that is involved in the control of infection by dengue and Zika viruses in mosquitoes. Preliminary analyses suggested that the loqs2 gene is a paralog of loquacious (loqs) and r2d2, two co-factors of the RNA interference (RNAi) pathway, a major antiviral mechanism in insects.
Results
Here we analyzed the origin and evolution of loqs2. Our data suggest that loqs2 originated from two independent duplications of the first double-stranded RNA binding domain of loqs that occurred before the origin of the Aedes Stegomyia subgenus, around 31 million years ago. We show that the loqs2 gene is evolving under relaxed purifying selection at a faster pace than loqs, with evidence of neofunctionalization driven by positive selection. Accordingly, we observed that Loqs2 is localized mainly in the nucleus, different from R2D2 and both isoforms of Loqs that are cytoplasmic. In contrast to r2d2 and loqs, loqs2 expression is stage- and tissue-specific, restricted mostly to reproductive tissues in adult Ae. aegypti and Ae. albopictus. Transgenic mosquitoes engineered to express loqs2 ubiquitously undergo developmental arrest at larval stages that correlates with massive dysregulation of gene expression without major effects on microRNAs or other endogenous small RNAs, classically associated with RNA interference.
Conclusions
Our results uncover the peculiar origin and neofunctionalization of loqs2 driven by positive selection. This study shows an example of unique adaptations in Aedes mosquitoes that could ultimately help explain their effectiveness as virus vectors.},
keywords = {Aedes mosquitoes, Double-stranded RNA (dsRNA), dsRNA binding protein (dsRBP), loqs2, M3i, marois, Marques, Olmo, RNA interference (RNAi)},
pubstate = {published},
tppubtype = {article}
}
Background
Mosquito borne viruses, such as dengue, Zika, yellow fever and Chikungunya, cause millions of infections every year. These viruses are mostly transmitted by two urban-adapted mosquito species, Aedes aegypti and Aedes albopictus. Although mechanistic understanding remains largely unknown, Aedes mosquitoes may have unique adaptations that lower the impact of viral infection. Recently, we reported the identification of an Aedes specific double-stranded RNA binding protein (dsRBP), named Loqs2, that is involved in the control of infection by dengue and Zika viruses in mosquitoes. Preliminary analyses suggested that the loqs2 gene is a paralog of loquacious (loqs) and r2d2, two co-factors of the RNA interference (RNAi) pathway, a major antiviral mechanism in insects.
Results
Here we analyzed the origin and evolution of loqs2. Our data suggest that loqs2 originated from two independent duplications of the first double-stranded RNA binding domain of loqs that occurred before the origin of the Aedes Stegomyia subgenus, around 31 million years ago. We show that the loqs2 gene is evolving under relaxed purifying selection at a faster pace than loqs, with evidence of neofunctionalization driven by positive selection. Accordingly, we observed that Loqs2 is localized mainly in the nucleus, different from R2D2 and both isoforms of Loqs that are cytoplasmic. In contrast to r2d2 and loqs, loqs2 expression is stage- and tissue-specific, restricted mostly to reproductive tissues in adult Ae. aegypti and Ae. albopictus. Transgenic mosquitoes engineered to express loqs2 ubiquitously undergo developmental arrest at larval stages that correlates with massive dysregulation of gene expression without major effects on microRNAs or other endogenous small RNAs, classically associated with RNA interference.
Conclusions
Our results uncover the peculiar origin and neofunctionalization of loqs2 driven by positive selection. This study shows an example of unique adaptations in Aedes mosquitoes that could ultimately help explain their effectiveness as virus vectors.
Mosquito borne viruses, such as dengue, Zika, yellow fever and Chikungunya, cause millions of infections every year. These viruses are mostly transmitted by two urban-adapted mosquito species, Aedes aegypti and Aedes albopictus. Although mechanistic understanding remains largely unknown, Aedes mosquitoes may have unique adaptations that lower the impact of viral infection. Recently, we reported the identification of an Aedes specific double-stranded RNA binding protein (dsRBP), named Loqs2, that is involved in the control of infection by dengue and Zika viruses in mosquitoes. Preliminary analyses suggested that the loqs2 gene is a paralog of loquacious (loqs) and r2d2, two co-factors of the RNA interference (RNAi) pathway, a major antiviral mechanism in insects.
Results
Here we analyzed the origin and evolution of loqs2. Our data suggest that loqs2 originated from two independent duplications of the first double-stranded RNA binding domain of loqs that occurred before the origin of the Aedes Stegomyia subgenus, around 31 million years ago. We show that the loqs2 gene is evolving under relaxed purifying selection at a faster pace than loqs, with evidence of neofunctionalization driven by positive selection. Accordingly, we observed that Loqs2 is localized mainly in the nucleus, different from R2D2 and both isoforms of Loqs that are cytoplasmic. In contrast to r2d2 and loqs, loqs2 expression is stage- and tissue-specific, restricted mostly to reproductive tissues in adult Ae. aegypti and Ae. albopictus. Transgenic mosquitoes engineered to express loqs2 ubiquitously undergo developmental arrest at larval stages that correlates with massive dysregulation of gene expression without major effects on microRNAs or other endogenous small RNAs, classically associated with RNA interference.
Conclusions
Our results uncover the peculiar origin and neofunctionalization of loqs2 driven by positive selection. This study shows an example of unique adaptations in Aedes mosquitoes that could ultimately help explain their effectiveness as virus vectors.
Green Emily I, Jaouen Etienne, Klug Dennis, Olmo Roenick Proveti, Gautier Amandine, Blandin Stéphanie, Marois Eric
A population modification gene drive targeting both Saglin and Lipophorin disables Plasmodium transmission in Anopheles mosquitoes Journal Article
In: Genetics and Genomics, 2024.
Abstract | Links | BibTeX | Tags: Anopheles, blandin, gene drive, Lipophorin, M3i, marois, marque, Marques, Olmo, Plasmodium, Saglin
@article{Green2024,
title = {A population modification gene drive targeting both Saglin and Lipophorin disables Plasmodium transmission in Anopheles mosquitoes},
author = {Emily I Green and Etienne Jaouen and Dennis Klug and Roenick Proveti Olmo and Amandine Gautier and Stéphanie Blandin and Eric Marois},
url = {https://elifesciences.org/articles/93142},
doi = {10.7554/eLife.93142},
year = {2024},
date = {2024-01-12},
journal = {Genetics and Genomics},
abstract = {Lipophorin is an essential, highly expressed lipid transport protein that is secreted and circulates in insect hemolymph. We hijacked the Anopheles coluzzii Lipophorin gene to make it co-express a single-chain version of antibody 2A10, which binds sporozoites of the malaria parasite Plasmodium falciparum. The resulting transgenic mosquitoes show a markedly decreased ability to transmit Plasmodium berghei expressing the P. falciparum circumsporozoite protein to mice. To force the spread of this antimalarial transgene in a mosquito population, we designed and tested several CRISPR/Cas9-based gene drives. One of these is installed in, and disrupts, the pro-parasitic gene Saglin and also cleaves wild-type Lipophorin, causing the anti-malarial modified Lipophorin version to replace the wild type and hitch-hike together with the Saglin drive. Although generating drive-resistant alleles and showing instability in its gRNA-encoding multiplex array, the Saglin-based gene drive reached high levels in caged mosquito populations and efficiently promoted the simultaneous spread of the antimalarial Lipophorin::Sc2A10 allele. This combination is expected to decrease parasite transmission via two different mechanisms. This work contributes to the design of novel strategies to spread antimalarial transgenes in mosquitoes, and illustrates some expected and unexpected outcomes encountered when establishing a population modification gene drive.},
keywords = {Anopheles, blandin, gene drive, Lipophorin, M3i, marois, marque, Marques, Olmo, Plasmodium, Saglin},
pubstate = {published},
tppubtype = {article}
}
Lipophorin is an essential, highly expressed lipid transport protein that is secreted and circulates in insect hemolymph. We hijacked the Anopheles coluzzii Lipophorin gene to make it co-express a single-chain version of antibody 2A10, which binds sporozoites of the malaria parasite Plasmodium falciparum. The resulting transgenic mosquitoes show a markedly decreased ability to transmit Plasmodium berghei expressing the P. falciparum circumsporozoite protein to mice. To force the spread of this antimalarial transgene in a mosquito population, we designed and tested several CRISPR/Cas9-based gene drives. One of these is installed in, and disrupts, the pro-parasitic gene Saglin and also cleaves wild-type Lipophorin, causing the anti-malarial modified Lipophorin version to replace the wild type and hitch-hike together with the Saglin drive. Although generating drive-resistant alleles and showing instability in its gRNA-encoding multiplex array, the Saglin-based gene drive reached high levels in caged mosquito populations and efficiently promoted the simultaneous spread of the antimalarial Lipophorin::Sc2A10 allele. This combination is expected to decrease parasite transmission via two different mechanisms. This work contributes to the design of novel strategies to spread antimalarial transgenes in mosquitoes, and illustrates some expected and unexpected outcomes encountered when establishing a population modification gene drive.
Marois Eric
Using the CRISPR / Cas9 system for genome editing in Anopheles mosquitoes Online
2024, visited: 08.01.2024.
Abstract | Links | BibTeX | Tags: Anopheles, CRISPR/Cas9, Genome editing, M3i, marois, mosquitoes
@online{Marois2024,
title = {Using the CRISPR / Cas9 system for genome editing in Anopheles mosquitoes},
author = { Eric Marois},
url = {https://hal.science/hal-04380430/document},
doi = {HAL Id: hal-04380430},
year = {2024},
date = {2024-01-08},
urldate = {2024-01-08},
journal = {HAL science},
abstract = {The advent of the CRISPR / Cas9 technology permits the targeted editing of mosquito genomes, ranging from site-directed mutagenesis of genes of interest yielding knockout mutations (which arise by insertion / deletion of a few nucleotides) to site-specific insertion of exogenous DNA sequences such as fluorescence markers or even large gene drive cassettes, themselves encoding the components of the CRISPR / Cas9 system. To obtain these heritable targeted changes, genome editing requires the delivery of Cas9 protein and its guide RNA(s) to the developing germ tissue of an embryo. Different species require adaptation of this basic principle to accommodate for their specific biology. Here, we describe a technical pipeline based on delivering the CRISPR/Cas9 components in the form of injected plasmid or as transgenes, resulting in highly efficient gene editing in Anopheles malaria vector mosquitoes. We have reliably employed these methods to mutagenize > 20 different loci of interest in Anopheles coluzzii to date. },
keywords = {Anopheles, CRISPR/Cas9, Genome editing, M3i, marois, mosquitoes},
pubstate = {published},
tppubtype = {online}
}
The advent of the CRISPR / Cas9 technology permits the targeted editing of mosquito genomes, ranging from site-directed mutagenesis of genes of interest yielding knockout mutations (which arise by insertion / deletion of a few nucleotides) to site-specific insertion of exogenous DNA sequences such as fluorescence markers or even large gene drive cassettes, themselves encoding the components of the CRISPR / Cas9 system. To obtain these heritable targeted changes, genome editing requires the delivery of Cas9 protein and its guide RNA(s) to the developing germ tissue of an embryo. Different species require adaptation of this basic principle to accommodate for their specific biology. Here, we describe a technical pipeline based on delivering the CRISPR/Cas9 components in the form of injected plasmid or as transgenes, resulting in highly efficient gene editing in Anopheles malaria vector mosquitoes. We have reliably employed these methods to mutagenize > 20 different loci of interest in Anopheles coluzzii to date.
2023
A Hinze, J Pelletier, M Ghaninia, E Marois, Hill, R S Ignell
Knockout of OR39 reveals redundancy in the olfactory pathway regulating the acquisition of host seeking in Anopheles coluzzii Journal Article
In: Proc Biol Sci 290(2011):20232092, vol. 290, no. 2011, 2023.
Abstract | Links | BibTeX | Tags: CRISPR–Cas9, human odour, M3i, marois, mosquito flight, odour valence, SSR
@article{Hinze2023,
title = {Knockout of OR39 reveals redundancy in the olfactory pathway regulating the acquisition of host seeking in Anopheles coluzzii},
author = {Hinze A and Pelletier J and Ghaninia M and Marois E and Hill and S Ignell R},
url = {https://doi.org/10.1098/rspb.2023.2092},
doi = {10.1098/rspb.2023.2092},
year = {2023},
date = {2023-11-29},
urldate = {2023-11-29},
booktitle = {Proc Biol Sci},
journal = {Proc Biol Sci 290(2011):20232092},
volume = {290},
number = {2011},
abstract = {The attraction of anthropophilic mosquitoes to human host cues, such as body odour and carbon dioxide, gradually increases during adult maturation. This acquisition of host-seeking behaviour correlates with age-dependent changes in odorant receptor (OR) transcript abundance and sensitivity of olfactory sensory neurons (OSNs). One OR gene of the human malaria vector, Anopheles coluzzii, AcolOR39, is significantly downregulated in mature females, and a cognate ligand of AcolOR39, sulcatone, a major component of human emanations, mediates the observed behavioural inhibition of newly emerged (teneral) females to human body odour. Knockout of AcolOR39, using CRISPR–Cas9 mutagenesis, selectively abolished sulcatone detection in OSNs, housed in trichoid sensilla. However, knockout of AcolOR39 altered neither the response rate nor the flight behaviour of teneral females in a wind tunnel, indicating the involvement of other genes, and thus a redundancy, in regulating the acquisition of host seeking in mosquitoes.},
keywords = {CRISPR–Cas9, human odour, M3i, marois, mosquito flight, odour valence, SSR},
pubstate = {published},
tppubtype = {article}
}
The attraction of anthropophilic mosquitoes to human host cues, such as body odour and carbon dioxide, gradually increases during adult maturation. This acquisition of host-seeking behaviour correlates with age-dependent changes in odorant receptor (OR) transcript abundance and sensitivity of olfactory sensory neurons (OSNs). One OR gene of the human malaria vector, Anopheles coluzzii, AcolOR39, is significantly downregulated in mature females, and a cognate ligand of AcolOR39, sulcatone, a major component of human emanations, mediates the observed behavioural inhibition of newly emerged (teneral) females to human body odour. Knockout of AcolOR39, using CRISPR–Cas9 mutagenesis, selectively abolished sulcatone detection in OSNs, housed in trichoid sensilla. However, knockout of AcolOR39 altered neither the response rate nor the flight behaviour of teneral females in a wind tunnel, indicating the involvement of other genes, and thus a redundancy, in regulating the acquisition of host seeking in mosquitoes.
SC Weng, I Antoshechkin, E Marois, O Akbari
Efficient Sex Separation by Exploiting Differential Alternative Splicing of a Dominant Marker in Aedes aegypti Journal Article
In: PLoS Genetics, 2023.
Abstract | Links | BibTeX | Tags: Aedes aegypti, Introns, larvae, M3i, marois, mosquitoes, pupae, rna sequencing, transcriptome analyses
@article{nokey,
title = {Efficient Sex Separation by Exploiting Differential Alternative Splicing of a Dominant Marker in Aedes aegypti},
author = {Weng SC and Antoshechkin I and Marois E and Akbari O},
url = {https://doi.org/10.1371/journal.pgen.1011065},
doi = {10.1371/journal.pgen.1011065},
year = {2023},
date = {2023-11-27},
urldate = {2023-11-27},
journal = {PLoS Genetics},
abstract = {Only female mosquitoes consume blood giving them the opportunity to transmit deadly human pathogens. Therefore, it is critical to remove females before conducting releases for genetic biocontrol interventions. Here we describe a robust sex-sorting approach termed SEPARATOR (Sexing Element Produced by Alternative RNA-splicing of A Transgenic Observable Reporter) that exploits sex-specific alternative splicing of an innocuous reporter to ensure exclusive dominant male-specific expression. Using SEPARATOR, we demonstrate reliable sex selection from early larval and pupal stages in Aedes aegypti, and use a Complex Object Parametric Analyzer and Sorter (COPAS) to demonstrate scalable high-throughput sex-selection of first instar larvae. Additionally, we use this approach to sequence the transcriptomes of early larval males and females and find several genes that are sex-specifically expressed. SEPARATOR can simplify mass production of males for release programs and is designed to be cross-species portable and should be instrumental for genetic biocontrol interventions.},
keywords = {Aedes aegypti, Introns, larvae, M3i, marois, mosquitoes, pupae, rna sequencing, transcriptome analyses},
pubstate = {published},
tppubtype = {article}
}
Only female mosquitoes consume blood giving them the opportunity to transmit deadly human pathogens. Therefore, it is critical to remove females before conducting releases for genetic biocontrol interventions. Here we describe a robust sex-sorting approach termed SEPARATOR (Sexing Element Produced by Alternative RNA-splicing of A Transgenic Observable Reporter) that exploits sex-specific alternative splicing of an innocuous reporter to ensure exclusive dominant male-specific expression. Using SEPARATOR, we demonstrate reliable sex selection from early larval and pupal stages in Aedes aegypti, and use a Complex Object Parametric Analyzer and Sorter (COPAS) to demonstrate scalable high-throughput sex-selection of first instar larvae. Additionally, we use this approach to sequence the transcriptomes of early larval males and females and find several genes that are sex-specifically expressed. SEPARATOR can simplify mass production of males for release programs and is designed to be cross-species portable and should be instrumental for genetic biocontrol interventions.
AI Kalita, E Marois, M Kozielska, FJ Weissing, E Jaouen, Möckel MM and Rühle F, F Butter, MF Basilicata, CI Keller Valsecchi
The sex-specific factor SOA controls dosage compensation in Anopheles mosquitoes Journal Article
In: Nature, vol. 623, pp. 175–182, 2023.
Abstract | Links | BibTeX | Tags: Anopheles, M3i, marois, SOA
@article{AI2023,
title = {The sex-specific factor SOA controls dosage compensation in Anopheles mosquitoes},
author = {Kalita AI and Marois E and Kozielska M and Weissing FJ and Jaouen E and Möckel MM and, Rühle F and Butter F and Basilicata MF and Keller Valsecchi CI},
url = {https://www.nature.com/articles/s41586-023-06641-0},
doi = {10.1038/s41586-023-06641-0.},
year = {2023},
date = {2023-09-28},
journal = {Nature},
volume = {623},
pages = {175–182},
abstract = {The Anopheles mosquito is one of thousands of species in which sex differences play a central part in their biology, as only females need a blood meal to produce eggs. Sex differentiation is regulated by sex chromosomes, but their presence creates a dosage imbalance between males (XY) and females (XX). Dosage compensation (DC) can re-equilibrate the expression of sex chromosomal genes. However, because DC mechanisms have only been fully characterized in a few model organisms, key questions about its evolutionary diversity and functional necessity remain unresolved1. Here we report the discovery of a previously uncharacterized gene (sex chromosome activation (SOA)) as a master regulator of DC in the malaria mosquito Anopheles gambiae. Sex-specific alternative splicing prevents functional SOA protein expression in females. The male isoform encodes a DNA-binding protein that binds the promoters of active X chromosomal genes. Expressing male SOA is sufficient to induce DC in female cells. Male mosquitoes lacking SOA or female mosquitoes ectopically expressing the male isoform exhibit X chromosome misregulation, which is compatible with viability but causes developmental delay. Thus, our molecular analyses of a DC master regulator in a non-model organism elucidates the evolutionary steps that lead to the establishment of a chromosome-specific fine-tuning mechanism.},
keywords = {Anopheles, M3i, marois, SOA},
pubstate = {published},
tppubtype = {article}
}
The Anopheles mosquito is one of thousands of species in which sex differences play a central part in their biology, as only females need a blood meal to produce eggs. Sex differentiation is regulated by sex chromosomes, but their presence creates a dosage imbalance between males (XY) and females (XX). Dosage compensation (DC) can re-equilibrate the expression of sex chromosomal genes. However, because DC mechanisms have only been fully characterized in a few model organisms, key questions about its evolutionary diversity and functional necessity remain unresolved1. Here we report the discovery of a previously uncharacterized gene (sex chromosome activation (SOA)) as a master regulator of DC in the malaria mosquito Anopheles gambiae. Sex-specific alternative splicing prevents functional SOA protein expression in females. The male isoform encodes a DNA-binding protein that binds the promoters of active X chromosomal genes. Expressing male SOA is sufficient to induce DC in female cells. Male mosquitoes lacking SOA or female mosquitoes ectopically expressing the male isoform exhibit X chromosome misregulation, which is compatible with viability but causes developmental delay. Thus, our molecular analyses of a DC master regulator in a non-model organism elucidates the evolutionary steps that lead to the establishment of a chromosome-specific fine-tuning mechanism.
SR Abbo, de Almeida JPP, RP Olmo, C Balvers, JS Griep, C Linthout, CJM Koenraadt, BM Silva, JJ Fros, ERGR Aguiar, E Marois, GP Pijlman, JT Marques
The virome of the invasive Asian bush mosquito Aedes japonicus in Europe Journal Article
In: Virus Evol., vol. 9, iss. 2, 2023.
Abstract | Links | BibTeX | Tags: Aedes japonicus, anphevirus, bunyavirus, M3i, marois, Marques, metagenomics, mosquito, Olmo, rhabdovirus, RNA Interference, totivirus, virome
@article{Abbo2023,
title = {The virome of the invasive Asian bush mosquito Aedes japonicus in Europe},
author = {Abbo SR and de Almeida JPP and Olmo RP and Balvers C and Griep JS and Linthout C and Koenraadt CJM and Silva BM and Fros JJ and Aguiar ERGR and Marois E and Pijlman GP and Marques JT},
doi = {10.1093/ve/vead041},
year = {2023},
date = {2023-09-22},
urldate = {2023-09-22},
journal = {Virus Evol.},
volume = {9},
issue = {2},
abstract = {The Asian bush mosquito Aedes japonicus is rapidly invading North America and Europe. Due to its potential to transmit multiple pathogenic arthropod-borne (arbo)viruses including Zika virus, West Nile virus, and chikungunya virus, it is important to understand the biology of this vector mosquito in more detail. In addition to arboviruses, mosquitoes can also carry insect-specific viruses that are receiving increasing attention due to their potential effects on host physiology and arbovirus transmission. In this study, we characterized the collection of viruses, referred to as the virome, circulating in Ae. japonicus populations in the Netherlands and France. Applying a small RNA-based metagenomic approach to Ae. japonicus, we uncovered a distinct group of viruses present in samples from both the Netherlands and France. These included one known virus, Ae. japonicus narnavirus 1 (AejapNV1), and three new virus species that we named Ae. japonicus totivirus 1 (AejapTV1), Ae. japonicus anphevirus 1 (AejapAV1) and Ae. japonicus bunyavirus 1 (AejapBV1). We also discovered sequences that were presumably derived from two additional novel viruses: Ae. japonicus bunyavirus 2 (AejapBV2) and Ae. japonicus rhabdovirus 1 (AejapRV1). All six viruses induced strong RNA interference responses, including the production of twenty-one nucleotide-sized small interfering RNAs, a signature of active replication in the host. Notably, AejapBV1 and AejapBV2 belong to different viral families; however, no RNA-dependent RNA polymerase sequence has been found for AejapBV2. Intriguingly, our small RNA-based approach identified an ∼1-kb long ambigrammatic RNA that is associated with AejapNV1 as a secondary segment but showed no similarity to any sequence in public databases. We confirmed the presence of AejapNV1 primary and secondary segments, AejapTV1, AejapAV1, and AejapBV1 by reverse transcriptase polymerase chain reaction (PCR) in wild-caught Ae. japonicus mosquitoes. AejapNV1 and AejapTV1 were found at high prevalence (87-100 per cent) in adult females, adult males, and larvae. Using a small RNA-based, sequence-independent metagenomic strategy, we uncovered a conserved and prevalent virome among Ae. japonicus mosquito populations. The high prevalence of AejapNV1 and AejapTV1 across all tested mosquito life stages suggests that these viruses are intimately associated with Ae. japonicus.},
keywords = {Aedes japonicus, anphevirus, bunyavirus, M3i, marois, Marques, metagenomics, mosquito, Olmo, rhabdovirus, RNA Interference, totivirus, virome},
pubstate = {published},
tppubtype = {article}
}
The Asian bush mosquito Aedes japonicus is rapidly invading North America and Europe. Due to its potential to transmit multiple pathogenic arthropod-borne (arbo)viruses including Zika virus, West Nile virus, and chikungunya virus, it is important to understand the biology of this vector mosquito in more detail. In addition to arboviruses, mosquitoes can also carry insect-specific viruses that are receiving increasing attention due to their potential effects on host physiology and arbovirus transmission. In this study, we characterized the collection of viruses, referred to as the virome, circulating in Ae. japonicus populations in the Netherlands and France. Applying a small RNA-based metagenomic approach to Ae. japonicus, we uncovered a distinct group of viruses present in samples from both the Netherlands and France. These included one known virus, Ae. japonicus narnavirus 1 (AejapNV1), and three new virus species that we named Ae. japonicus totivirus 1 (AejapTV1), Ae. japonicus anphevirus 1 (AejapAV1) and Ae. japonicus bunyavirus 1 (AejapBV1). We also discovered sequences that were presumably derived from two additional novel viruses: Ae. japonicus bunyavirus 2 (AejapBV2) and Ae. japonicus rhabdovirus 1 (AejapRV1). All six viruses induced strong RNA interference responses, including the production of twenty-one nucleotide-sized small interfering RNAs, a signature of active replication in the host. Notably, AejapBV1 and AejapBV2 belong to different viral families; however, no RNA-dependent RNA polymerase sequence has been found for AejapBV2. Intriguingly, our small RNA-based approach identified an ∼1-kb long ambigrammatic RNA that is associated with AejapNV1 as a secondary segment but showed no similarity to any sequence in public databases. We confirmed the presence of AejapNV1 primary and secondary segments, AejapTV1, AejapAV1, and AejapBV1 by reverse transcriptase polymerase chain reaction (PCR) in wild-caught Ae. japonicus mosquitoes. AejapNV1 and AejapTV1 were found at high prevalence (87-100 per cent) in adult females, adult males, and larvae. Using a small RNA-based, sequence-independent metagenomic strategy, we uncovered a conserved and prevalent virome among Ae. japonicus mosquito populations. The high prevalence of AejapNV1 and AejapTV1 across all tested mosquito life stages suggests that these viruses are intimately associated with Ae. japonicus.
Marois Eric
Screening Mosquito Larvae Under a Fluorescence Binocular Microscope Journal Article
In: Cold Spring Harb Protoc, 2023.
Links | BibTeX | Tags: fluorescent protein, M3i, marois
@article{Marois2023,
title = {Screening Mosquito Larvae Under a Fluorescence Binocular Microscope},
author = {Eric Marois},
doi = {10.1101/pdb.prot108306 },
year = {2023},
date = {2023-09-11},
urldate = {2023-09-11},
journal = {Cold Spring Harb Protoc},
keywords = {fluorescent protein, M3i, marois},
pubstate = {published},
tppubtype = {article}
}
PELLETIER Julien, DAWIT Mengistu, GHANINIA Majid, MAROIS Eric, IGNELL Rickard
A mosquito-specific antennal protein is critical for the attraction to human odor in the malaria vector Anopheles gambiae Journal Article
In: Insect Biochemistry and Molecular Biology, vol. 159, iss. August 2023, 2023.
Abstract | Links | BibTeX | Tags: antenna, chemoreceptor, M3i, marois, mosquitoes, olfaction
@article{IGNELL2023,
title = {A mosquito-specific antennal protein is critical for the attraction to human odor in the malaria vector Anopheles gambiae},
author = {Julien PELLETIER AND Mengistu DAWIT AND Majid GHANINIA AND Eric MAROIS AND Rickard IGNELL},
editor = { },
url = {https://doi.org/10.1016/j.ibmb.2023.103988},
doi = {j.ibmb.2023.103988},
year = {2023},
date = {2023-07-11},
urldate = {2023-07-11},
journal = {Insect Biochemistry and Molecular Biology},
volume = {159},
issue = {August 2023},
abstract = {Mosquitoes rely mainly on the sense of smell to decipher their environment and locate suitable food sources, hosts for blood feeding and oviposition sites. The molecular bases of olfaction involve multigenic families of olfactory proteins that have evolved to interact with a narrow set of odorants that are critical for survival. Understanding the complex interplay between diversified repertoires of olfactory proteins and ecologically-relevant odorant signals, which elicit important behaviors, is fundamental for the design of novel control strategies targeting the sense of smell of disease vector mosquitoes. Previously, large multigene families of odorant receptor and ionotropic receptor proteins, as well as a subset of odorant-binding proteins have been shown to mediate the selectivity and sensitivity of the mosquito olfactory system. In this study, we identify a mosquito-specific antennal protein (MSAP) gene as a novel molecular actor of odorant reception. MSAP is highly conserved across mosquito species and is transcribed at an extremely high level in female antennae. In order to understand its role in the mosquito olfactory system, we generated knockout mutant lines in Anopheles gambiae, and performed comparative analysis of behavioral and physiological responses to human-associated odorants. We found that MSAP promotes female mosquito attraction to human odor and enhances the sensitivity of the antennae to a variety of odorants. These findings suggest that MSAP is an important component of the mosquito olfactory system, which until now has gone completely unnoticed.},
keywords = {antenna, chemoreceptor, M3i, marois, mosquitoes, olfaction},
pubstate = {published},
tppubtype = {article}
}
Mosquitoes rely mainly on the sense of smell to decipher their environment and locate suitable food sources, hosts for blood feeding and oviposition sites. The molecular bases of olfaction involve multigenic families of olfactory proteins that have evolved to interact with a narrow set of odorants that are critical for survival. Understanding the complex interplay between diversified repertoires of olfactory proteins and ecologically-relevant odorant signals, which elicit important behaviors, is fundamental for the design of novel control strategies targeting the sense of smell of disease vector mosquitoes. Previously, large multigene families of odorant receptor and ionotropic receptor proteins, as well as a subset of odorant-binding proteins have been shown to mediate the selectivity and sensitivity of the mosquito olfactory system. In this study, we identify a mosquito-specific antennal protein (MSAP) gene as a novel molecular actor of odorant reception. MSAP is highly conserved across mosquito species and is transcribed at an extremely high level in female antennae. In order to understand its role in the mosquito olfactory system, we generated knockout mutant lines in Anopheles gambiae, and performed comparative analysis of behavioral and physiological responses to human-associated odorants. We found that MSAP promotes female mosquito attraction to human odor and enhances the sensitivity of the antennae to a variety of odorants. These findings suggest that MSAP is an important component of the mosquito olfactory system, which until now has gone completely unnoticed.
C Lutrat, M Burckbuchler, RP Olmo, R Beugnon, A Fontaine, OS Akbari, R Argiles-Herrero, T Baldet, J Bouyer, E Marois
Combining two Genetic Sexing Strains allows sorting of non-transgenic males for Aedes genetic control Journal Article
In: Commun Biol., vol. 6, iss. 1, pp. 646, 2023.
Abstract | Links | BibTeX | Tags: Aedes, M3i, marois, mosquitoes, Olmo, vectoring
@article{Lutrat2023,
title = {Combining two Genetic Sexing Strains allows sorting of non-transgenic males for Aedes genetic control},
author = {Lutrat C and Burckbuchler M and Olmo RP and Beugnon R and Fontaine A and Akbari OS and Argiles-Herrero R and Baldet T and Bouyer J and Marois E},
url = {https://www.nature.com/articles/s42003-023-05030-7},
doi = {10.1038/s42003-023-05030-7},
year = {2023},
date = {2023-06-16},
urldate = {2023-06-16},
journal = {Commun Biol.},
volume = {6},
issue = {1},
pages = {646},
abstract = {Chemical control of disease vectoring mosquitoes Aedes albopictus and Aedes aegypti is costly, unsustainable, and increasingly ineffective due to the spread of insecticide resistance. The Sterile Insect Technique is a valuable alternative but is limited by slow, error-prone, and wasteful sex-separation methods. Here, we present four Genetic Sexing Strains (two for each Aedes species) based on fluorescence markers linked to the m and M sex loci, allowing for the isolation of transgenic males. Furthermore, we demonstrate how combining these sexing strains enables the production of non-transgenic males. In a mass-rearing facility, 100,000 first instar male larvae could be sorted in under 1.5 h with an estimated 0.01–0.1% female contamination on a single machine. Cost-efficiency analyses revealed that using these strains could result in important savings while setting up and running a mass-rearing facility. Altogether, these Genetic Sexing Strains should enable a major upscaling in control programmes against these important vectors.},
keywords = {Aedes, M3i, marois, mosquitoes, Olmo, vectoring},
pubstate = {published},
tppubtype = {article}
}
Chemical control of disease vectoring mosquitoes Aedes albopictus and Aedes aegypti is costly, unsustainable, and increasingly ineffective due to the spread of insecticide resistance. The Sterile Insect Technique is a valuable alternative but is limited by slow, error-prone, and wasteful sex-separation methods. Here, we present four Genetic Sexing Strains (two for each Aedes species) based on fluorescence markers linked to the m and M sex loci, allowing for the isolation of transgenic males. Furthermore, we demonstrate how combining these sexing strains enables the production of non-transgenic males. In a mass-rearing facility, 100,000 first instar male larvae could be sorted in under 1.5 h with an estimated 0.01–0.1% female contamination on a single machine. Cost-efficiency analyses revealed that using these strains could result in important savings while setting up and running a mass-rearing facility. Altogether, these Genetic Sexing Strains should enable a major upscaling in control programmes against these important vectors.
Klug Dennis, Gautier Amandine, Calvo Eric, Marois Eric, Blandin Stéphanie A.
The salivary protein Saglin facilitates efficient midgut colonization of Anopheles mosquitoes by malaria parasites Journal Article
In: Plos Pathogens, vol. 19, iss. 3, no. 3, 2023.
Abstract | Links | BibTeX | Tags: blandin, BLOOD, M3i, malarial parasites, marois, mosquitoes, Oocysts, Parasitic Diseases, Plasmodium, salivary glands, sporozoites
@article{Klug2023,
title = {The salivary protein Saglin facilitates efficient midgut colonization of Anopheles mosquitoes by malaria parasites},
author = {Dennis Klug and Amandine Gautier and Eric Calvo and Eric Marois and Stéphanie A. Blandin},
url = {https://doi.org/10.1371/journal.ppat.1010538},
doi = {10.1371/journal.ppat.1010538},
year = {2023},
date = {2023-03-02},
urldate = {2023-03-02},
booktitle = {Plos Pathogens},
journal = {Plos Pathogens},
volume = {19},
number = {3},
issue = {3},
abstract = {Female mosquitoes rely on blood feeding to acquire sufficient nutrients for egg development. Because of the importance of this process mosquitoes evolved salivary proteins with a broad range of functions acting as blood thinners, anti-coagulants and immunosuppressants. The effect of these proteins on the blood at the bite site directly influences the size of the blood bolus a female takes up in a given time frame. Both, time of feeding and bolus size, are important parameters for fecundity and survival. Recent studies have shown that a significant proportion of salivated proteins is re-ingested during feeding and becomes part of the blood meal. Here we investigated the salivary protein Saglin which has been previously suggested as putative receptor mediating malaria parasite entry into the salivary gland. By engineering a loss-of-function mutant in An. coluzzi we could show that the absence of Saglin impairs the development of parasite stages in the blood meal of the rodent malaria parasite P. berghei and the human malaria parasite P. falciparum lowering the parasite burden of subsequent stages and preventing efficient transmission at low infection densities. Furthermore, we could show that Saglin is present in the blood meal after feeding possibly indicating a previously overlooked parasite-vector interaction.},
keywords = {blandin, BLOOD, M3i, malarial parasites, marois, mosquitoes, Oocysts, Parasitic Diseases, Plasmodium, salivary glands, sporozoites},
pubstate = {published},
tppubtype = {article}
}
Female mosquitoes rely on blood feeding to acquire sufficient nutrients for egg development. Because of the importance of this process mosquitoes evolved salivary proteins with a broad range of functions acting as blood thinners, anti-coagulants and immunosuppressants. The effect of these proteins on the blood at the bite site directly influences the size of the blood bolus a female takes up in a given time frame. Both, time of feeding and bolus size, are important parameters for fecundity and survival. Recent studies have shown that a significant proportion of salivated proteins is re-ingested during feeding and becomes part of the blood meal. Here we investigated the salivary protein Saglin which has been previously suggested as putative receptor mediating malaria parasite entry into the salivary gland. By engineering a loss-of-function mutant in An. coluzzi we could show that the absence of Saglin impairs the development of parasite stages in the blood meal of the rodent malaria parasite P. berghei and the human malaria parasite P. falciparum lowering the parasite burden of subsequent stages and preventing efficient transmission at low infection densities. Furthermore, we could show that Saglin is present in the blood meal after feeding possibly indicating a previously overlooked parasite-vector interaction.
R.P. Olmo, Y.M.H. Todjro, E.R.G.R. Aguiar, de Almeida J.P.P., F.V. Ferreira, J.N. Armache, de Faria I.J.S., A.G.A. Ferreira, S.C.D. Amadou, A.T.S. Silva, de Souza K.P.R., A.P.P. Vilela, A. Babarit, C.H. Tan, M. Diallo, A. Gaye, C. Paupy, J. Obame-Nkoghe, T.M. Visser, C.J.M. Koenraadt, M.A. Wongsokarijo, A.L.C. Cruz, M.T. Prieto, M.C.P. Parra, Nogueira M.L., V. Avelino-Silva, R.N. Mota, M.A.Z. Borges, B.P. Drumond, E.G. Kroon, M. Recker, L. Sedda, E. Marois, J.L. Imler, J.T. Marques
Mosquito vector competence for dengue is modulated by insect-specific viruses Journal Article
In: Nature Microbiology, vol. 8, iss. 1, 2023.
Abstract | Links | BibTeX | Tags: Dengue, imler, M3i, marois, Marques, mosquito, Olmo, virus
@article{Olmo.2023,
title = {Mosquito vector competence for dengue is modulated by insect-specific viruses},
author = {Olmo R.P. and Todjro Y.M.H. and Aguiar E.R.G.R. and de Almeida J.P.P. and Ferreira F.V. and Armache J.N. and de Faria I.J.S. and Ferreira A.G.A. and Amadou S.C.D. and Silva A.T.S. and de Souza K.P.R. and Vilela A.P.P. and Babarit A. and Tan C.H. and Diallo M. and Gaye A. and Paupy C. and Obame-Nkoghe J. and Visser T.M. and Koenraadt C.J.M. and Wongsokarijo M.A. and Cruz A.L.C. and Prieto M.T. and Parra M.C.P. and Nogueira M.L., and Avelino-Silva V. and Mota R.N. and Borges M.A.Z. and Drumond B.P. and Kroon E.G. and Recker M. and Sedda L. and Marois E. and Imler J.L. and Marques J.T. },
url = {https://doi.org/},
doi = {10.1038/s41564-022-01289-4},
year = {2023},
date = {2023-01-05},
urldate = {2023-01-05},
journal = {Nature Microbiology},
volume = {8},
issue = {1},
abstract = {Aedes aegypti and A. albopictus mosquitoes are the main vectors for dengue virus (DENV) and other arboviruses, including Zika virus (ZIKV). Understanding the factors that affect transmission of arboviruses from mosquitoes to humans is a priority because it could inform public health and targeted interventions. Reasoning that interactions among viruses in the vector insect might affect transmission, we analysed the viromes of 815 urban Aedes mosquitoes collected from 12 countries worldwide. Two mosquito-specific viruses, Phasi Charoen-like virus (PCLV) and Humaita Tubiacanga virus (HTV), were the most abundant in A. aegypti worldwide. Spatiotemporal analyses of virus circulation in an endemic urban area revealed a 200% increase in chances of having DENV in wild A. aegypti mosquitoes when both HTV and PCLV were present. Using a mouse model in the laboratory, we showed that the presence of HTV and PCLV increased the ability of mosquitoes to transmit DENV and ZIKV to a vertebrate host. By transcriptomic analysis, we found that in DENV-infected mosquitoes, HTV and PCLV block the downregulation of histone H4, which we identify as an important proviral host factor in vivo.},
keywords = {Dengue, imler, M3i, marois, Marques, mosquito, Olmo, virus},
pubstate = {published},
tppubtype = {article}
}
Aedes aegypti and A. albopictus mosquitoes are the main vectors for dengue virus (DENV) and other arboviruses, including Zika virus (ZIKV). Understanding the factors that affect transmission of arboviruses from mosquitoes to humans is a priority because it could inform public health and targeted interventions. Reasoning that interactions among viruses in the vector insect might affect transmission, we analysed the viromes of 815 urban Aedes mosquitoes collected from 12 countries worldwide. Two mosquito-specific viruses, Phasi Charoen-like virus (PCLV) and Humaita Tubiacanga virus (HTV), were the most abundant in A. aegypti worldwide. Spatiotemporal analyses of virus circulation in an endemic urban area revealed a 200% increase in chances of having DENV in wild A. aegypti mosquitoes when both HTV and PCLV were present. Using a mouse model in the laboratory, we showed that the presence of HTV and PCLV increased the ability of mosquitoes to transmit DENV and ZIKV to a vertebrate host. By transcriptomic analysis, we found that in DENV-infected mosquitoes, HTV and PCLV block the downregulation of histone H4, which we identify as an important proviral host factor in vivo.
2020
Volohonsky Gloria, Paul-Gilloteaux Perrine, Stafkova Jitka, Soichot Julien, Salamero Jean, Levashina Elena A.
Kinetics of Plasmodium midgut invasion in Anopheles mosquitoes Journal Article
In: PLoS Pathog, vol. 16, no. 9, 2020, ISSN: 1553-7374.
Abstract | Links | BibTeX | Tags: Anopheles, M3i, Malaria, marois, midgut, Plasmodium
@article{volohonsky_plasmodium_2020,
title = {Kinetics of Plasmodium midgut invasion in Anopheles mosquitoes},
author = {Gloria Volohonsky and Perrine Paul-Gilloteaux and Jitka Stafkova and Julien Soichot and Jean Salamero and Elena A. Levashina
},
editor = {Kenneth D. Vernick, Institut Pasteur, FRANCE},
url = {https://journals.plos.org/plospathogens/article?id=10.1371/journal.ppat.1008739},
doi = {10.1371/journal.ppat.1008739},
issn = {1553-7374},
year = {2020},
date = {2020-09-18},
journal = {PLoS Pathog},
volume = {16},
number = {9},
abstract = {The traversal of the mosquito midgut cells is one of the critical stages in the life cycle of malaria parasites. Motile parasite forms, called ookinetes, traverse the midgut epithelium in a dynamic process which is not fully understood. Here, we harnessed transgenic reporters to track invasion of Plasmodium parasites in African and Indian mosquito species. We found important differences in parasite dynam- ics between the two Anopheles species and demonstrated a role of the mosquito comple- ment-like system in regulation of parasite invasion of the midgut cells.},
keywords = {Anopheles, M3i, Malaria, marois, midgut, Plasmodium},
pubstate = {published},
tppubtype = {article}
}
The traversal of the mosquito midgut cells is one of the critical stages in the life cycle of malaria parasites. Motile parasite forms, called ookinetes, traverse the midgut epithelium in a dynamic process which is not fully understood. Here, we harnessed transgenic reporters to track invasion of Plasmodium parasites in African and Indian mosquito species. We found important differences in parasite dynam- ics between the two Anopheles species and demonstrated a role of the mosquito comple- ment-like system in regulation of parasite invasion of the midgut cells.
2019
Achee Nicole L., Grieco John P., Vatandoost Hassan, Seixas Gonçalo, Pinto Joao, Ching-NG Lee, Martins Ademir J., Juntarajumnong Waraporn, Corbel Vincent, Gouagna Clement, David Jean-Philippe, Logan James G., Orsborne James, Marois Eric, Devine Gregor J., Vontas John
Alternative strategies for mosquito-borne arbovirus control Journal Article
In: PLoS Neglected Tropical Diseases, 2019.
Abstract | Links | BibTeX | Tags: arbovirus, M3i, marois, mosquito
@article{Achee2019,
title = {Alternative strategies for mosquito-borne arbovirus control},
author = { Nicole L. Achee and John P. Grieco and Hassan Vatandoost and Gonçalo Seixas and Joao Pinto and Lee Ching-NG and Ademir J. Martins and Waraporn Juntarajumnong and Vincent Corbel and Clement Gouagna and Jean-Philippe David and James G. Logan and James Orsborne and Eric Marois and Gregor J. Devine and John Vontas
},
url = {https://doi.org/10.1371/journal.pntd.0006822},
doi = {10.1371/journal.pntd.0006822},
year = {2019},
date = {2019-01-03},
journal = {PLoS Neglected Tropical Diseases},
abstract = {International public health workers are challenged by the burden of arthropod-borne viral diseases, to include mosquito-borne arboviruses transmitted by Aedes aegypti and A. albopictus due in part to lack of sustainable vector control and insecticide resistance (IR), as well as the inability to scale up and sustain existing interventions for prevention of urban epidemics. As a consequence, there has been increasing interest to advance the development of alternative methods. This review provides a general overview of alternative vector-control strategies under development for the control of arbovirus mosquito vectors and highlights how each could offer innovative public health value. Considerations to regulations, acceptance, and sustainability are also provided.},
keywords = {arbovirus, M3i, marois, mosquito},
pubstate = {published},
tppubtype = {article}
}
International public health workers are challenged by the burden of arthropod-borne viral diseases, to include mosquito-borne arboviruses transmitted by Aedes aegypti and A. albopictus due in part to lack of sustainable vector control and insecticide resistance (IR), as well as the inability to scale up and sustain existing interventions for prevention of urban epidemics. As a consequence, there has been increasing interest to advance the development of alternative methods. This review provides a general overview of alternative vector-control strategies under development for the control of arbovirus mosquito vectors and highlights how each could offer innovative public health value. Considerations to regulations, acceptance, and sustainability are also provided.
2018
Olmo RP, Ferreira AGA, Izidoro-Toledo TC, Aguiar ERGR, de Faria IJS, de Souza KPR, Osório KP, Kuhn L, Hammann P, de Andrade EG, Todjro YM, Rocha MN, Leite THJF, Amadou SCG, Armache JN, Paro S, de Oliveira CD, Carvalho FD, Moreira LA, Marois E, Imler JL, Marques JT
Control of dengue virus in the midgut of Aedes aegypti by ectopic expression of the dsRNA-binding protein Loqs2 Journal Article
In: Nature Microbiology, vol. 3, no. 12, pp. 1385-1393, 2018.
Abstract | Links | BibTeX | Tags: Aedes aegypti, Dengue, imler, M3i, marois, Marques, Zika
@article{Olmo_2018,
title = {Control of dengue virus in the midgut of Aedes aegypti by ectopic expression of the dsRNA-binding protein Loqs2 },
author = {RP Olmo and AGA Ferreira and TC Izidoro-Toledo and ERGR Aguiar and IJS de Faria and KPR de Souza and KP Osório and L Kuhn and P Hammann and EG de Andrade and YM Todjro and MN Rocha and THJF Leite and SCG Amadou and JN Armache and S Paro and CD de Oliveira and FD Carvalho and LA Moreira and E Marois and JL Imler and JT Marques},
url = {https://www.nature.com/articles/s41564-018-0268-6},
doi = {10.1038/s41564-018-0268-6},
year = {2018},
date = {2018-10-29},
journal = {Nature Microbiology},
volume = {3},
number = {12},
pages = {1385-1393},
abstract = {Dengue virus (DENV) is an arbovirus transmitted to humans by Aedes mosquitoes. In the insect vector, the small interfering RNA (siRNA) pathway is an important antiviral mechanism against DENV. However, it remains unclear when and where the siRNA pathway acts during the virus cycle. Here, we show that the siRNA pathway fails to efficiently silence DENV in the midgut of Aedes aegypti although it is essential to restrict systemic replication. Accumulation of DENV-derived siRNAs in the midgut reveals that impaired silencing results from a defect downstream of small RNA biogenesis. Notably, silencing triggered by endogenous and exogenous dsRNAs remained effective in the midgut where known components of the siRNA pathway, including the double-stranded RNA (dsRNA)-binding proteins Loquacious and r2d2, had normal expression levels. We identified an Aedes-specific paralogue of loquacious and r2d2, hereafter named loqs2, which is not expressed in the midgut. Loqs2 interacts with Loquacious and r2d2 and is required to control systemic replication of DENV and also Zika virus. Furthermore, ectopic expression of Loqs2 in the midgut of transgenic mosquitoes is sufficient to restrict DENV replication and dissemination. Together, our data reveal a mechanism of tissue-specific regulation of the mosquito siRNA pathway controlled by Loqs2. },
keywords = {Aedes aegypti, Dengue, imler, M3i, marois, Marques, Zika},
pubstate = {published},
tppubtype = {article}
}
Dengue virus (DENV) is an arbovirus transmitted to humans by Aedes mosquitoes. In the insect vector, the small interfering RNA (siRNA) pathway is an important antiviral mechanism against DENV. However, it remains unclear when and where the siRNA pathway acts during the virus cycle. Here, we show that the siRNA pathway fails to efficiently silence DENV in the midgut of Aedes aegypti although it is essential to restrict systemic replication. Accumulation of DENV-derived siRNAs in the midgut reveals that impaired silencing results from a defect downstream of small RNA biogenesis. Notably, silencing triggered by endogenous and exogenous dsRNAs remained effective in the midgut where known components of the siRNA pathway, including the double-stranded RNA (dsRNA)-binding proteins Loquacious and r2d2, had normal expression levels. We identified an Aedes-specific paralogue of loquacious and r2d2, hereafter named loqs2, which is not expressed in the midgut. Loqs2 interacts with Loquacious and r2d2 and is required to control systemic replication of DENV and also Zika virus. Furthermore, ectopic expression of Loqs2 in the midgut of transgenic mosquitoes is sufficient to restrict DENV replication and dissemination. Together, our data reveal a mechanism of tissue-specific regulation of the mosquito siRNA pathway controlled by Loqs2.
J. Bouyer, E. Marois
Pests and vector-borne diseases in the livestock industry Book Chapter
In: Academic, Wageningen (Ed.): vol. 5, Chapter Genetic control of vectors, pp. 435–451, 2018, ISBN: 9789086868636 .
Abstract | Links | BibTeX | Tags: insect resistance, M3i, marois
@inbook{Bouyer2018,
title = {Pests and vector-borne diseases in the livestock industry},
author = {Bouyer J. and Marois E.},
editor = {Wageningen Academic},
url = {https://doi.org/10.3920/978-90-8686-863-6_14 },
doi = {10.3920/978-90-8686-863-6_14},
isbn = {9789086868636 },
year = {2018},
date = {2018-08-15},
volume = {5},
pages = {435–451},
chapter = {Genetic control of vectors},
series = {Ecology and Control of Vector-borne Diseases},
abstract = {In a context of tighter regulations on approved insecticide molecules, the spread of insecticide resistance in insect vectors of human and animal diseases and the introduction of exotic vectors to new territories call for the development of new pest control methods and strategies. New genetic control methods, related to the ancestral sterile insect technique (SIT), show particular promise and are being developed in response to increasing health and agricultural challenges. These include the use of symbionts like Wolbachia and the use of transgenic insect strains, some of which incorporate gene editing techniques that can lead to transgene spread (gene drive). Here we present the principles, associated opportunities and risks, as well as the degree of advancement of these various techniques for a subset of livestock pests and disease vectors including screwworms, tsetse, mosquitoes and stomoxes. We then present some case studies on recent improvements in the use of the SIT in tsetse and the release of insects carrying a dominant lethal gene, symbiont-based approaches and gene drive in mosquitoes. Finally, we call to speed up the development of genetic control, within a rigorous benefit-risk analysis framework including international public consultation.},
keywords = {insect resistance, M3i, marois},
pubstate = {published},
tppubtype = {inbook}
}
In a context of tighter regulations on approved insecticide molecules, the spread of insecticide resistance in insect vectors of human and animal diseases and the introduction of exotic vectors to new territories call for the development of new pest control methods and strategies. New genetic control methods, related to the ancestral sterile insect technique (SIT), show particular promise and are being developed in response to increasing health and agricultural challenges. These include the use of symbionts like Wolbachia and the use of transgenic insect strains, some of which incorporate gene editing techniques that can lead to transgene spread (gene drive). Here we present the principles, associated opportunities and risks, as well as the degree of advancement of these various techniques for a subset of livestock pests and disease vectors including screwworms, tsetse, mosquitoes and stomoxes. We then present some case studies on recent improvements in the use of the SIT in tsetse and the release of insects carrying a dominant lethal gene, symbiont-based approaches and gene drive in mosquitoes. Finally, we call to speed up the development of genetic control, within a rigorous benefit-risk analysis framework including international public consultation.
Dong Yuemei, Simões Maria L., Marois Eric, Dimopoulos George
CRISPR/Cas9 -mediated gene knockout of Anopheles gambiae FREP1 suppresses malaria parasite infection Journal Article
In: PLoS Pathog, vol. 14, no. 3, 2018.
Abstract | Links | BibTeX | Tags: Anopheles gambiae, CRISPR/Cas9, Knockout, M3i, Malaria, marois
@article{Dong2018,
title = {CRISPR/Cas9 -mediated gene knockout of Anopheles gambiae FREP1 suppresses malaria parasite infection},
author = { Yuemei Dong and Maria L. Simões and Eric Marois and George Dimopoulos },
url = {https://doi.org/10.1371/journal.ppat.1006898},
doi = {10.1371/journal.ppat.1006898},
year = {2018},
date = {2018-03-08},
urldate = {2018-03-08},
journal = {PLoS Pathog},
volume = {14},
number = {3},
abstract = {Plasmodium relies on numerous agonists during its journey through the mosquito vector, and these agonists represent potent targets for transmission-blocking by either inhibiting or interfering with them pre- or post-transcriptionally. The recently developed CRISPR/Cas9-based genome editing tools for Anopheles mosquitoes provide new and promising opportunities for the study of agonist function and for developing malaria control strategies through gene deletion to achieve complete agonist inactivation. Here we have established a modified CRISPR/Cas9 gene editing procedure for the malaria vector Anopheles gambiae, and studied the effect of inactivating the fibrinogen-related protein 1 (FREP1) gene on the mosquito’s susceptibility to Plasmodium and on mosquito fitness. FREP1 knockout mutants developed into adult mosquitoes that showed profound suppression of infection with both human and rodent malaria parasites at the oocyst and sporozoite stages. FREP1 inactivation, however, resulted in fitness costs including a significantly lower blood-feeding propensity, fecundity and egg hatching rate, a retarded pupation time, and reduced longevity after a blood meal.},
keywords = {Anopheles gambiae, CRISPR/Cas9, Knockout, M3i, Malaria, marois},
pubstate = {published},
tppubtype = {article}
}
Plasmodium relies on numerous agonists during its journey through the mosquito vector, and these agonists represent potent targets for transmission-blocking by either inhibiting or interfering with them pre- or post-transcriptionally. The recently developed CRISPR/Cas9-based genome editing tools for Anopheles mosquitoes provide new and promising opportunities for the study of agonist function and for developing malaria control strategies through gene deletion to achieve complete agonist inactivation. Here we have established a modified CRISPR/Cas9 gene editing procedure for the malaria vector Anopheles gambiae, and studied the effect of inactivating the fibrinogen-related protein 1 (FREP1) gene on the mosquito’s susceptibility to Plasmodium and on mosquito fitness. FREP1 knockout mutants developed into adult mosquitoes that showed profound suppression of infection with both human and rodent malaria parasites at the oocyst and sporozoite stages. FREP1 inactivation, however, resulted in fitness costs including a significantly lower blood-feeding propensity, fecundity and egg hatching rate, a retarded pupation time, and reduced longevity after a blood meal.
2017
G Triller, SW Scally, G Costa, M Pissarev, C Kreschel, A Bosch, E Marois, BK Sack, R Murugan, AM Salman, CJ Janse, SM Khan, SH Kappe, AA Adegnika, B Mordmüller, EA Levashina, JP Julien, H Wardemann
Natural parasite exposure induces protective human anti-malarial antibodies Journal Article
In: Immunity , vol. 47, iss. 6, pp. 1197-1209, 2017.
Abstract | Links | BibTeX | Tags: anti-malarial antibodie, M3i, marois
@article{Triller2017,
title = {Natural parasite exposure induces protective human anti-malarial antibodies},
author = {Triller G and Scally SW and Costa G and Pissarev M and Kreschel C and Bosch A and Marois E and Sack BK and Murugan R and Salman AM and Janse CJ and Khan SM and Kappe SH and Adegnika AA and Mordmüller B and Levashina EA and Julien JP and Wardemann H},
url = {https://doi.org/10.1016/j.immuni.2017.11.007},
doi = {10.1016/j.immuni.2017.11.007},
year = {2017},
date = {2017-12-19},
journal = {Immunity },
volume = {47},
issue = {6},
pages = {1197-1209},
abstract = {Antibodies against the NANP repeat of circumsporozoite protein (CSP), the major surface antigen of Plasmodium falciparum (Pf) sporozoites, can protect from malaria in animal models but protective humoral immunity is difficult to induce in humans. Here we cloned and characterized rare affinity-matured human NANP-reactive memory B cell antibodies elicited by natural Pf exposure that potently inhibited parasite transmission and development in vivo. We unveiled the molecular details of antibody binding to two distinct protective epitopes within the NANP repeat. NANP repeat recognition was largely mediated by germline encoded and immunoglobulin (Ig) heavy-chain complementarity determining region 3 (HCDR3) residues, whereas affinity maturation contributed predominantly to stabilizing the antigen-binding site conformation. Combined, our findings illustrate the power of exploring human anti-CSP antibody responses to develop tools for malaria control in the mammalian and the mosquito vector and provide a molecular basis for the structure-based design of next-generation CSP malaria vaccines. },
keywords = {anti-malarial antibodie, M3i, marois},
pubstate = {published},
tppubtype = {article}
}
Antibodies against the NANP repeat of circumsporozoite protein (CSP), the major surface antigen of Plasmodium falciparum (Pf) sporozoites, can protect from malaria in animal models but protective humoral immunity is difficult to induce in humans. Here we cloned and characterized rare affinity-matured human NANP-reactive memory B cell antibodies elicited by natural Pf exposure that potently inhibited parasite transmission and development in vivo. We unveiled the molecular details of antibody binding to two distinct protective epitopes within the NANP repeat. NANP repeat recognition was largely mediated by germline encoded and immunoglobulin (Ig) heavy-chain complementarity determining region 3 (HCDR3) residues, whereas affinity maturation contributed predominantly to stabilizing the antigen-binding site conformation. Combined, our findings illustrate the power of exploring human anti-CSP antibody responses to develop tools for malaria control in the mammalian and the mosquito vector and provide a molecular basis for the structure-based design of next-generation CSP malaria vaccines.
Volohonsky Gloria, Hopp Ann-Katrin, Saenger Mélanie, Soichot Julien, Scholze Heidi, Boch Jens, Blandin Stéphanie A, Marois Eric
Transgenic Expression of the Anti-parasitic Factor TEP1 in the Malaria Mosquito Anopheles gambiae Journal Article
In: PLOS Pathogens, vol. 13, no. 1, pp. e1006113, 2017, ISSN: 1553-7374.
Links | BibTeX | Tags: Anopheles gambiae, anti-parasitic factor, blandin, M3i, Malaria, marois, TEP1, transgenic
@article{volohonsky_transgenic_2017,
title = {Transgenic Expression of the Anti-parasitic Factor TEP1 in the Malaria Mosquito Anopheles gambiae},
author = {Gloria Volohonsky and Ann-Katrin Hopp and Mélanie Saenger and Julien Soichot and Heidi Scholze and Jens Boch and Stéphanie A Blandin and Eric Marois},
editor = {Kenneth D Vernick},
url = {http://dx.plos.org/10.1371/journal.ppat.1006113},
doi = {10.1371/journal.ppat.1006113},
issn = {1553-7374},
year = {2017},
date = {2017-01-01},
urldate = {2017-02-01},
journal = {PLOS Pathogens},
volume = {13},
number = {1},
pages = {e1006113},
keywords = {Anopheles gambiae, anti-parasitic factor, blandin, M3i, Malaria, marois, TEP1, transgenic},
pubstate = {published},
tppubtype = {article}
}
2016
Ehrhardt Katharina, Deregnaucourt Christiane, Goetz Alice-Anne, Tzanova Tzvetomira, Gallo Valentina, Arese Paolo, Pradines Bruno, Adjalley Sophie H, Bagrel Denyse, Blandin Stephanie A, Lanzer Michael, Davioud-Charvet Elisabeth
In: Antimicrob. Agents Chemother., vol. 60, no. 9, pp. 5146-5158, 2016, ISSN: 1098-6596.
Abstract | Links | BibTeX | Tags: antimalarial, blandin, M3i, parasites, Plasmodium, redox-cycler
@article{ehrhardt_redox-cycler_2016,
title = {The redox-cycler plasmodione is a fast acting antimalarial lead compound with pronounced activity against sexual and early asexual blood-stage parasites},
author = {Katharina Ehrhardt and Christiane Deregnaucourt and Alice-Anne Goetz and Tzvetomira Tzanova and Valentina Gallo and Paolo Arese and Bruno Pradines and Sophie H Adjalley and Denyse Bagrel and Stephanie A Blandin and Michael Lanzer and Elisabeth Davioud-Charvet},
url = {http://aac.asm.org/content/60/9/5146},
doi = {10.1128/AAC.02975-15},
issn = {1098-6596},
year = {2016},
date = {2016-09-01},
journal = {Antimicrob. Agents Chemother.},
volume = {60},
number = {9},
pages = {5146-5158},
abstract = {Previously, we presented the chemical design of a promising series of antimalarial agents, 3-[substituted-benzyl]-menadiones, with potent in vitro and in vivo activities. Ongoing studies on the mode of action of antimalarial 3-[substituted-benzyl]-menadiones revealed that these agents disturb the redox balance of the parasitized erythrocyte by acting as redox-cyclers - a strategy that is broadly recognized for the development of new antimalarial agents. Here, we report a detailed parasitological characterization of the in vitro activity profile of the lead compound 3-[4-(trifluoromethyl)benzyl]-menadione 1c (henceforth called plasmodione) against intra-erythrocytic stages of the human malaria parasite Plasmodium falciparum We show that plasmodione acts rapidly against asexual blood stages, thereby disrupting the clinically relevant intra-erythrocytic life cycle of the parasite, and furthermore has potent activity against early gametocytes. The lead's antiplasmodial activity was unaffected by the most common resistance mechanisms to clinically used antimalarials. Moreover, plasmodione has a low potential to induce drug resistance and a fast killing speed as observed by culturing parasites under continuous drug pressure. Drug interactions with licensed antimalarial drugs were also established using the fixed-ratio isobologram method. Initial toxicological profiling suggests that it is a safe agent for possible human use. Our studies identify plasmodione as a promising antimalarial lead compound and strongly support the future development of redox-active benzylmenadiones as antimalarial agents.},
keywords = {antimalarial, blandin, M3i, parasites, Plasmodium, redox-cycler},
pubstate = {published},
tppubtype = {article}
}
Previously, we presented the chemical design of a promising series of antimalarial agents, 3-[substituted-benzyl]-menadiones, with potent in vitro and in vivo activities. Ongoing studies on the mode of action of antimalarial 3-[substituted-benzyl]-menadiones revealed that these agents disturb the redox balance of the parasitized erythrocyte by acting as redox-cyclers - a strategy that is broadly recognized for the development of new antimalarial agents. Here, we report a detailed parasitological characterization of the in vitro activity profile of the lead compound 3-[4-(trifluoromethyl)benzyl]-menadione 1c (henceforth called plasmodione) against intra-erythrocytic stages of the human malaria parasite Plasmodium falciparum We show that plasmodione acts rapidly against asexual blood stages, thereby disrupting the clinically relevant intra-erythrocytic life cycle of the parasite, and furthermore has potent activity against early gametocytes. The lead's antiplasmodial activity was unaffected by the most common resistance mechanisms to clinically used antimalarials. Moreover, plasmodione has a low potential to induce drug resistance and a fast killing speed as observed by culturing parasites under continuous drug pressure. Drug interactions with licensed antimalarial drugs were also established using the fixed-ratio isobologram method. Initial toxicological profiling suggests that it is a safe agent for possible human use. Our studies identify plasmodione as a promising antimalarial lead compound and strongly support the future development of redox-active benzylmenadiones as antimalarial agents.
2015
Volohonsky Gloria, Terenzi Olivier, Soichot Julien, Naujoks Daniel A, Nolan Tony, Windbichler Nikolai, Kapps Delphine, Smidler Andie L, Vittu Anaïs, Costa Giulia, Steinert Stefanie, Levashina Elena A, Blandin Stéphanie A, Marois Eric
Tools for Anopheles gambiae Transgenesis Journal Article
In: G3 (Bethesda), vol. 5, no. 6, pp. 1151-63, 2015.
Abstract | Links | BibTeX | Tags: Anopheles gambiae, bioinformatic, blandin, M3i, marois, transgenesis
@article{G2015,
title = {Tools for Anopheles gambiae Transgenesis},
author = {Gloria Volohonsky and Olivier Terenzi and Julien Soichot and Daniel A Naujoks and Tony Nolan and Nikolai Windbichler and Delphine Kapps and Andie L Smidler and Anaïs Vittu and Giulia Costa and Stefanie Steinert and Elena A Levashina and Stéphanie A Blandin and Eric Marois},
url = {http://www.ncbi.nlm.nih.gov/pubmed/25869647},
year = {2015},
date = {2015-04-13},
journal = {G3 (Bethesda)},
volume = {5},
number = {6},
pages = {1151-63},
abstract = {Transgenesis is an essential tool to investigate gene function and to introduce desired characters in laboratory organisms. Setting-up transgenesis in non-model organisms is challenging due to the diversity of biological life traits and due to knowledge gaps in genomic information. Some procedures will be broadly applicable to many organisms, and others have to be specifically developed for the target species. Transgenesis in disease vector mosquitoes has existed since the 2000s but has remained limited by the delicate biology of these insects. Here, we report a compilation of the transgenesis tools that we have designed for the malaria vector Anopheles gambiae, including new docking strains, convenient transgenesis plasmids, a puromycin resistance selection marker, mosquitoes expressing cre recombinase, and various reporter lines defining the activity of cloned promoters. This toolbox contributed to rendering transgenesis routine in this species and is now enabling the development of increasingly refined genetic manipulations such as targeted mutagenesis. Some of the reagents and procedures reported here are easily transferable to other nonmodel species, including other disease vector or agricultural pest insects.},
keywords = {Anopheles gambiae, bioinformatic, blandin, M3i, marois, transgenesis},
pubstate = {published},
tppubtype = {article}
}
Transgenesis is an essential tool to investigate gene function and to introduce desired characters in laboratory organisms. Setting-up transgenesis in non-model organisms is challenging due to the diversity of biological life traits and due to knowledge gaps in genomic information. Some procedures will be broadly applicable to many organisms, and others have to be specifically developed for the target species. Transgenesis in disease vector mosquitoes has existed since the 2000s but has remained limited by the delicate biology of these insects. Here, we report a compilation of the transgenesis tools that we have designed for the malaria vector Anopheles gambiae, including new docking strains, convenient transgenesis plasmids, a puromycin resistance selection marker, mosquitoes expressing cre recombinase, and various reporter lines defining the activity of cloned promoters. This toolbox contributed to rendering transgenesis routine in this species and is now enabling the development of increasingly refined genetic manipulations such as targeted mutagenesis. Some of the reagents and procedures reported here are easily transferable to other nonmodel species, including other disease vector or agricultural pest insects.
Neafsey D E, Waterhouse R M, Abai M R, Aganezov S S, Alekseyev M A, Allen J E, Amon J, Arcà B, Arensburger P, Artemov G, Assour L A, Basseri H, Berlin A, Birren B W, Blandin Stéphanie A, Brockman A I, Burkot T R, Burt A, Chan C S, Chauve C, Chiu J C, Christensen M, Costantini C, Davidson V L M, Deligianni E, Dottorini T, Dritsou V, Gabriel S B, Guelbeogo W M, Hall A B, Han M W, Hlaing T, Hughes D S T, Jenkins A M, Jiang X, Jungreis I, Kakani E G, Kamali M, Kemppainen P, Kennedy R C, Kirmitzoglou I K, Koekemoer L L, Laban N, Langridge N, Lawniczak M K N, Lirakis M, Lobo N F, Lowy E, MacCallum R M, Mao C, Maslen G, Mbogo C, McCarthy J, Michel K, Mitchell S N, Moore W, Murphy K A, Naumenko A N, Nolan Tony, Novoa E M, O’Loughlin S, Oringanje C, Oshaghi M A, Pakpour N, Papathanos P A, Peery A N, Povelones Michael, Prakash A, Price D A, Rajaraman A, Reimer L J, Rinker D C, Rokas A, Russell T L, Sagnon N F, Sharakhova M V, Shea T, Simão F A, Simard F, Slotman M A, Somboon P, Stegniy V, Struchiner C J, Thomas G W C, Tojo M, Topalis P, Tubio J M C, Unger M F, Vontas J, Walton C, Wilding C S, Willis J H, Wu Y-C, Yan G, Zdobnov E M, Zhou X, Catteruccia Flaminia, Christophides Georges K, Collins F H, Cornman R S, Crisanti Andrea, Donnelly M J, Emrich S J, Fontaine M C, Gelbart W, Hahn M W, Hansen I A, Howell P I, Kafatos Fotis C, Kellis M, Lawson D, Louis C, Luckhart S, Muskavitch M A T, Ribeiro J M, Riehle M A, Sharakhov I V, Tu Z, Zwiebel L J, Besansky N J
Highly evolvable malaria vectors: The genomes of 16 Anopheles mosquitoes. Journal Article
In: Science, vol. 347, no. 6217, pp. 1258522, 2015.
Abstract | Links | BibTeX | Tags: Anopheles, blandin, genomes, M3i
@article{DE2015,
title = {Highly evolvable malaria vectors: The genomes of 16 Anopheles mosquitoes.},
author = {D E Neafsey and R M Waterhouse and M R Abai and S S Aganezov and M A Alekseyev and J E Allen and J Amon and B Arcà and P Arensburger and G Artemov and L A Assour and H Basseri and A Berlin and B W Birren and Stéphanie A Blandin and A I Brockman and T R Burkot and A Burt and C S Chan and C Chauve and J C Chiu and M Christensen and C Costantini and V L M Davidson and E Deligianni and T Dottorini and V Dritsou and S B Gabriel and W M Guelbeogo and A B Hall and M W Han and T Hlaing and D S T Hughes and A M Jenkins and X Jiang and I Jungreis and E G Kakani and M Kamali and P Kemppainen and R C Kennedy and I K Kirmitzoglou and L L Koekemoer and N Laban and N Langridge and M K N Lawniczak and M Lirakis and N F Lobo and E Lowy and R M MacCallum and C Mao and G Maslen and C Mbogo and J McCarthy and K Michel and S N Mitchell and W Moore and K A Murphy and A N Naumenko and Tony Nolan and E M Novoa and S O’Loughlin and C Oringanje and M A Oshaghi and N Pakpour and P A Papathanos and A N Peery and Michael Povelones and A Prakash and D A Price and A Rajaraman and L J Reimer and D C Rinker and A Rokas and T L Russell and N F Sagnon and M V Sharakhova and T Shea and F A Simão and F Simard and M A Slotman and P Somboon and V Stegniy and C J Struchiner and G W C Thomas and M Tojo and P Topalis and J M C Tubio and M F Unger and J Vontas and C Walton and C S Wilding and J H Willis and Y-C Wu and G Yan and E M Zdobnov and X Zhou and Flaminia Catteruccia and Georges K Christophides and F H Collins and R S Cornman and Andrea Crisanti and M J Donnelly and S J Emrich and M C Fontaine and W Gelbart and M W Hahn and I A Hansen and P I Howell and Fotis C Kafatos and M Kellis and D Lawson and C Louis and S Luckhart and M A T Muskavitch and J M Ribeiro and M A Riehle and I V Sharakhov and Z Tu and L J Zwiebel and N J Besansky},
url = {http://www.ncbi.nlm.nih.gov/pubmed/25554792},
year = {2015},
date = {2015-01-02},
journal = {Science},
volume = {347},
number = {6217},
pages = {1258522},
abstract = {Variation in vectorial capacity for human malaria among Anopheles mosquito species is determined by many factors, including behavior, immunity, and life history. To investigate the genomic basis of vectorial capacity and explore new avenues for vector control, we sequenced the genomes of 16 anopheline mosquito species from diverse locations spanning ~100 million years of evolution. Comparative analyses show faster rates of gene gain and loss, elevated gene shuffling on the X chromosome, and more intron losses, relative to Drosophila. Some determinants of vectorial capacity, such as chemosensory genes, do not show elevated turnover but instead diversify through protein-sequence changes. This dynamism of anopheline genes and genomes may contribute to their flexible capacity to take advantage of new ecological niches, including adapting to humans as primary hosts.},
keywords = {Anopheles, blandin, genomes, M3i},
pubstate = {published},
tppubtype = {article}
}
Variation in vectorial capacity for human malaria among Anopheles mosquito species is determined by many factors, including behavior, immunity, and life history. To investigate the genomic basis of vectorial capacity and explore new avenues for vector control, we sequenced the genomes of 16 anopheline mosquito species from diverse locations spanning ~100 million years of evolution. Comparative analyses show faster rates of gene gain and loss, elevated gene shuffling on the X chromosome, and more intron losses, relative to Drosophila. Some determinants of vectorial capacity, such as chemosensory genes, do not show elevated turnover but instead diversify through protein-sequence changes. This dynamism of anopheline genes and genomes may contribute to their flexible capacity to take advantage of new ecological niches, including adapting to humans as primary hosts.
Hammond Andrew, Galizi Roberto, Kyrou Kyros, Simoni Alekos, Siniscalchi Carla, Katsanos Dimitris, Gribble Matthew, Baker Dean, Marois Eric, Russell Steven, Burt Austin, Windbichler Nikolai, Crisanti Andrea, Nolan Tony
A CRISPR-Cas9 gene drive system targeting female reproduction in the malaria mosquito vector Anopheles gambiae Journal Article
In: Nature Biotechnology, vol. 34, no. 1, pp. 78–83, 2015, ISSN: 1087-0156, 1546-1696.
Links | BibTeX | Tags: M3i, marois
@article{hammond_crispr-cas9_2015,
title = {A CRISPR-Cas9 gene drive system targeting female reproduction in the malaria mosquito vector Anopheles gambiae},
author = {Andrew Hammond and Roberto Galizi and Kyros Kyrou and Alekos Simoni and Carla Siniscalchi and Dimitris Katsanos and Matthew Gribble and Dean Baker and Eric Marois and Steven Russell and Austin Burt and Nikolai Windbichler and Andrea Crisanti and Tony Nolan},
url = {http://www.nature.com/doifinder/10.1038/nbt.3439},
doi = {10.1038/nbt.3439},
issn = {1087-0156, 1546-1696},
year = {2015},
date = {2015-01-01},
urldate = {2016-01-26},
journal = {Nature Biotechnology},
volume = {34},
number = {1},
pages = {78--83},
keywords = {M3i, marois},
pubstate = {published},
tppubtype = {article}
}
2014
Gabrieli P, Marois Eric, Catteruccia Flaminia
Transgenic insects: techniques and applications Book Chapter
In: Benedicts, MQ (Ed.): Chapter Sexual sterilization of mosquitoes, pp. 188-207, CABI, 2014.
BibTeX | Tags: insects, M3i, marois, sterile
@inbook{P2014,
title = {Transgenic insects: techniques and applications},
author = {P Gabrieli and Eric Marois and Flaminia Catteruccia},
editor = {MQ Benedicts},
year = {2014},
date = {2014-10-01},
pages = {188-207},
publisher = {CABI},
chapter = {Sexual sterilization of mosquitoes},
keywords = {insects, M3i, marois, sterile},
pubstate = {published},
tppubtype = {inbook}
}
Bernardini F, Galizi R, Menichelli M, Papathanos P A, Dritsou V, Marois Eric, Crisanti Andrea, Windbichler Nikolai
Site-specific genetic engineering of the Anopheles gambiae Y chromosome Journal Article
In: Proc Natl Acad Sci U S A., vol. 111, no. 21, pp. 7600-5, 2014.
Abstract | Links | BibTeX | Tags: Anopheles gambiae, Biotechnology, M3i, marois, SIT, transgenesis
@article{F2014b,
title = {Site-specific genetic engineering of the Anopheles gambiae Y chromosome},
author = {F Bernardini and R Galizi and M Menichelli and P A Papathanos and V Dritsou and Eric Marois and Andrea Crisanti and Nikolai Windbichler},
url = {http://www.ncbi.nlm.nih.gov/pubmed/24821795},
year = {2014},
date = {2014-05-27},
journal = {Proc Natl Acad Sci U S A.},
volume = {111},
number = {21},
pages = {7600-5},
abstract = {Despite its function in sex determination and its role in driving genome evolution, the Y chromosome remains poorly understood in most species. Y chromosomes are gene-poor, repeat-rich and largely heterochromatic and therefore represent a difficult target for genetic engineering. The Y chromosome of the human malaria vector Anopheles gambiae appears to be involved in sex determination although very little is known about both its structure and function. Here, we characterize a transgenic strain of this mosquito species, obtained by transposon-mediated integration of a transgene construct onto the Y chromosome. Using meganuclease-induced homologous repair we introduce a site-specific recombination signal onto the Y chromosome and show that the resulting docking line can be used for secondary integration. To demonstrate its utility, we study the activity of a germ-line-specific promoter when located on the Y chromosome. We also show that Y-linked fluorescent transgenes allow automated sex separation of this important vector species, providing the means to generate large single-sex populations. Our findings will aid studies of sex chromosome function and enable the development of male-exclusive genetic traits for vector control.},
keywords = {Anopheles gambiae, Biotechnology, M3i, marois, SIT, transgenesis},
pubstate = {published},
tppubtype = {article}
}
Despite its function in sex determination and its role in driving genome evolution, the Y chromosome remains poorly understood in most species. Y chromosomes are gene-poor, repeat-rich and largely heterochromatic and therefore represent a difficult target for genetic engineering. The Y chromosome of the human malaria vector Anopheles gambiae appears to be involved in sex determination although very little is known about both its structure and function. Here, we characterize a transgenic strain of this mosquito species, obtained by transposon-mediated integration of a transgene construct onto the Y chromosome. Using meganuclease-induced homologous repair we introduce a site-specific recombination signal onto the Y chromosome and show that the resulting docking line can be used for secondary integration. To demonstrate its utility, we study the activity of a germ-line-specific promoter when located on the Y chromosome. We also show that Y-linked fluorescent transgenes allow automated sex separation of this important vector species, providing the means to generate large single-sex populations. Our findings will aid studies of sex chromosome function and enable the development of male-exclusive genetic traits for vector control.
2013
Smidler Andie L, Terenzi Olivier, Soichot Julien, Levashina Elena A, Marois Eric
Targeted Mutagenesis in the Malaria Mosquito Using TALE Nucleases Journal Article
In: PLoS One, vol. 8, no. 8, pp. e74511, 2013.
Abstract | Links | BibTeX | Tags: M3i, marois, Mutagenesis, TALEN
@article{AL2013,
title = {Targeted Mutagenesis in the Malaria Mosquito Using TALE Nucleases},
author = {Andie L Smidler and Olivier Terenzi and Julien Soichot and Elena A Levashina and Eric Marois},
url = {http://www.ncbi.nlm.nih.gov/pubmed/23977401},
year = {2013},
date = {2013-08-15},
journal = {PLoS One},
volume = {8},
number = {8},
pages = {e74511},
abstract = {Anopheles gambiae, the main mosquito vector of human malaria, is a challenging organism to manipulate genetically. As a consequence, reverse genetics studies in this disease vector have been largely limited to RNA interference experiments. Here, we report the targeted disruption of the immunity gene TEP1 using transgenic expression of Transcription-Activator Like Effector Nucleases (TALENs), and the isolation of several TEP1 mutant A. gambiae lines. These mutations inhibited protein production and rendered TEP1 mutants hypersusceptible to Plasmodium berghei. The TALEN technology opens up new avenues for genetic analysis in this disease vector and may offer novel biotechnology-based approaches for malaria control.},
keywords = {M3i, marois, Mutagenesis, TALEN},
pubstate = {published},
tppubtype = {article}
}
Anopheles gambiae, the main mosquito vector of human malaria, is a challenging organism to manipulate genetically. As a consequence, reverse genetics studies in this disease vector have been largely limited to RNA interference experiments. Here, we report the targeted disruption of the immunity gene TEP1 using transgenic expression of Transcription-Activator Like Effector Nucleases (TALENs), and the isolation of several TEP1 mutant A. gambiae lines. These mutations inhibited protein production and rendered TEP1 mutants hypersusceptible to Plasmodium berghei. The TALEN technology opens up new avenues for genetic analysis in this disease vector and may offer novel biotechnology-based approaches for malaria control.
Lamacchia Marina, Clayton John Randy, Wang-Sattler R, Steinmetz Lars M, Levashina Elena A, Blandin Stéphanie A
Silencing of Genes and Alleles by RNAi in Anopheles gambiae Journal Article
In: Methods Mol Biol., vol. 923, pp. 161-76, 2013.
Abstract | BibTeX | Tags: blandin, dsRNA, M3i, RNAi
@article{M2013,
title = {Silencing of Genes and Alleles by RNAi in Anopheles gambiae},
author = {Marina Lamacchia and John Randy Clayton and R Wang-Sattler and Lars M Steinmetz and Elena A Levashina and Stéphanie A Blandin},
year = {2013},
date = {2013-06-13},
journal = {Methods Mol Biol.},
volume = {923},
pages = {161-76},
abstract = {Anopheles gambiae mosquitoes are the major vectors of human malaria parasites. However, mosquitoes are not passive hosts for parasites, actively limiting their development in vivo. Our current understanding of the mosquito antiparasitic response is mostly based on the phenotypic analysis of gene knockdowns obtained by RNA interference (RNAi), through the injection or transfection of long dsRNAs in adult mosquitoes or cultured cells, respectively. Recently, RNAi has been extended to silence specifically one allele of a given gene in a heterozygous context, thus allowing to compare the contribution of different alleles to a phenotype in the same genetic background.},
keywords = {blandin, dsRNA, M3i, RNAi},
pubstate = {published},
tppubtype = {article}
}
Anopheles gambiae mosquitoes are the major vectors of human malaria parasites. However, mosquitoes are not passive hosts for parasites, actively limiting their development in vivo. Our current understanding of the mosquito antiparasitic response is mostly based on the phenotypic analysis of gene knockdowns obtained by RNA interference (RNAi), through the injection or transfection of long dsRNAs in adult mosquitoes or cultured cells, respectively. Recently, RNAi has been extended to silence specifically one allele of a given gene in a heterozygous context, thus allowing to compare the contribution of different alleles to a phenotype in the same genetic background.
2012
Marois Eric, Scali C, Soichot Julien, Kappler Christine, Levashina Elena A, Catteruccia Flaminia
High-throughput sorting of mosquito larvae for laboratory studies and for future vector control interventions Journal Article
In: Malaria J., vol. 11, pp. 302, 2012.
Abstract | Links | BibTeX | Tags: COPAS, M3i, marois, Sorting, transgenesis
@article{E2012,
title = {High-throughput sorting of mosquito larvae for laboratory studies and for future vector control interventions},
author = {Eric Marois and C Scali and Julien Soichot and Christine Kappler and Elena A Levashina and Flaminia Catteruccia},
url = {http://www.ncbi.nlm.nih.gov/pubmed/22929810},
year = {2012},
date = {2012-08-28},
journal = {Malaria J.},
volume = {11},
pages = {302},
abstract = {BACKGROUND: Mosquito transgenesis offers new promises for the genetic control of vector-borne infectious diseases such as malaria and dengue fever. Genetic control strategies require the release of large number of male mosquitoes into field populations, whether they are based on the use of sterile males (sterile insect technique, SIT) or on introducing genetic traits conferring refractoriness to disease transmission (population replacement). However, the current absence of high-throughput techniques for sorting different mosquito populations impairs the application of these control measures. METHODS: A method was developed to generate large mosquito populations of the desired sex and genotype. This method combines flow cytometry and the use of Anopheles gambiae transgenic lines that differentially express fluorescent markers in males and females. RESULTS: Fluorescence-assisted sorting allowed single-step isolation of homozygous transgenic mosquitoes from a mixed population. This method was also used to select wild-type males only with high efficiency and accuracy, a highly desirable tool for genetic control strategies where the release of transgenic individuals may be problematic. Importantly, sorted males showed normal mating ability compared to their unsorted brothers. CONCLUSIONS: The developed method will greatly facilitate both laboratory studies of mosquito vectorial capacity requiring high-throughput approaches and future field interventions in the fight against infectious disease vectors.},
keywords = {COPAS, M3i, marois, Sorting, transgenesis},
pubstate = {published},
tppubtype = {article}
}
BACKGROUND: Mosquito transgenesis offers new promises for the genetic control of vector-borne infectious diseases such as malaria and dengue fever. Genetic control strategies require the release of large number of male mosquitoes into field populations, whether they are based on the use of sterile males (sterile insect technique, SIT) or on introducing genetic traits conferring refractoriness to disease transmission (population replacement). However, the current absence of high-throughput techniques for sorting different mosquito populations impairs the application of these control measures. METHODS: A method was developed to generate large mosquito populations of the desired sex and genotype. This method combines flow cytometry and the use of Anopheles gambiae transgenic lines that differentially express fluorescent markers in males and females. RESULTS: Fluorescence-assisted sorting allowed single-step isolation of homozygous transgenic mosquitoes from a mixed population. This method was also used to select wild-type males only with high efficiency and accuracy, a highly desirable tool for genetic control strategies where the release of transgenic individuals may be problematic. Importantly, sorted males showed normal mating ability compared to their unsorted brothers. CONCLUSIONS: The developed method will greatly facilitate both laboratory studies of mosquito vectorial capacity requiring high-throughput approaches and future field interventions in the fight against infectious disease vectors.
2011
Marois Eric
The multifaceted mosquito anti-Plasmodium response Journal Article
In: Curr Opin Microbiol., vol. 14, no. 4, pp. 429-35, 2011.
Abstract | Links | BibTeX | Tags: anti-Plasmodium response, M3i, marois
@article{E2011,
title = {The multifaceted mosquito anti-Plasmodium response},
author = {Eric Marois},
url = {http://www.ncbi.nlm.nih.gov/pubmed/21802348},
year = {2011},
date = {2011-07-27},
journal = {Curr Opin Microbiol.},
volume = {14},
number = {4},
pages = {429-35},
abstract = {Plasmodium development within its mosquito vector is an essential step in malaria transmission, as illustrated in world regions where malaria was successfully eradicated via vector control. The innate immune system of most mosquitoes is able to completely clear a Plasmodium infection, preventing parasite transmission to humans. Understanding the biological basis of this phenomenon is expected to inspire new strategies to curb malaria incidence in countries where vector control via insecticides is unpractical, or inefficient because insecticide resistance genes have spread across mosquito populations. Several aspects of mosquito biology that condition the success of the parasite in colonizing its vector begin to be understood at the molecular level, and a wealth of recently published data highlights the multifaceted nature of the mosquito response against parasite invasion. In this brief review, we attempt to provide an integrated view of the challenges faced by the parasite to successfully invade its mosquito host, and discuss the possible intervention strategies that could exploit this knowledge for the fight against human malaria.},
keywords = {anti-Plasmodium response, M3i, marois},
pubstate = {published},
tppubtype = {article}
}
Plasmodium development within its mosquito vector is an essential step in malaria transmission, as illustrated in world regions where malaria was successfully eradicated via vector control. The innate immune system of most mosquitoes is able to completely clear a Plasmodium infection, preventing parasite transmission to humans. Understanding the biological basis of this phenomenon is expected to inspire new strategies to curb malaria incidence in countries where vector control via insecticides is unpractical, or inefficient because insecticide resistance genes have spread across mosquito populations. Several aspects of mosquito biology that condition the success of the parasite in colonizing its vector begin to be understood at the molecular level, and a wealth of recently published data highlights the multifaceted nature of the mosquito response against parasite invasion. In this brief review, we attempt to provide an integrated view of the challenges faced by the parasite to successfully invade its mosquito host, and discuss the possible intervention strategies that could exploit this knowledge for the fight against human malaria.
2010
Rono Martin K, Whitten Miranda M, Oulad-Abdelghani M, Levashina Elena A, Marois Eric
The Major Yolk Protein Vitellogenin Interferes with the Anti-Plasmodium Response in the Malaria Mosquito Anopheles gambiae Journal Article
In: PLoS Biol., vol. 10, no. 7, pp. e1000434, 2010.
Abstract | Links | BibTeX | Tags: anti-Plasmodium response, M3i, marois, vittelogenin
@article{MK2010,
title = {The Major Yolk Protein Vitellogenin Interferes with the Anti-Plasmodium Response in the Malaria Mosquito Anopheles gambiae},
author = {Martin K Rono and Miranda M Whitten and M Oulad-Abdelghani and Elena A Levashina and Eric Marois},
url = {http://www.ncbi.nlm.nih.gov/pubmed/20652016},
year = {2010},
date = {2010-07-20},
journal = {PLoS Biol.},
volume = {10},
number = {7},
pages = {e1000434},
abstract = {When taking a blood meal on a person infected with malaria, female Anopheles gambiae mosquitoes, the major vector of human malaria, acquire nutrients that will activate egg development (oogenesis) in their ovaries. Simultaneously, they infect themselves with the malaria parasite. On traversing the mosquito midgut epithelium, invading Plasmodium ookinetes are met with a potent innate immune response predominantly controlled by mosquito blood cells. Whether the concomitant processes of mosquito reproduction and immunity affect each other remains controversial. Here, we show that proteins that deliver nutrients to maturing mosquito oocytes interfere with the antiparasitic response. Lipophorin (Lp) and vitellogenin (Vg), two nutrient transport proteins, reduce the parasite-killing efficiency of the antiparasitic factor TEP1. In the absence of either nutrient transport protein, TEP1 binding to the ookinete surface becomes more efficient. We also show that Lp is required for the normal expression of Vg, and for later Plasmodium development at the oocyst stage. Furthermore, our results uncover an inhibitory role of the Cactus/REL1/REL2 signaling cassette in the expression of Vg, but not of Lp. We reveal molecular links that connect reproduction and immunity at several levels and provide a molecular basis for a long-suspected trade-off between these two processes.},
keywords = {anti-Plasmodium response, M3i, marois, vittelogenin},
pubstate = {published},
tppubtype = {article}
}
When taking a blood meal on a person infected with malaria, female Anopheles gambiae mosquitoes, the major vector of human malaria, acquire nutrients that will activate egg development (oogenesis) in their ovaries. Simultaneously, they infect themselves with the malaria parasite. On traversing the mosquito midgut epithelium, invading Plasmodium ookinetes are met with a potent innate immune response predominantly controlled by mosquito blood cells. Whether the concomitant processes of mosquito reproduction and immunity affect each other remains controversial. Here, we show that proteins that deliver nutrients to maturing mosquito oocytes interfere with the antiparasitic response. Lipophorin (Lp) and vitellogenin (Vg), two nutrient transport proteins, reduce the parasite-killing efficiency of the antiparasitic factor TEP1. In the absence of either nutrient transport protein, TEP1 binding to the ookinete surface becomes more efficient. We also show that Lp is required for the normal expression of Vg, and for later Plasmodium development at the oocyst stage. Furthermore, our results uncover an inhibitory role of the Cactus/REL1/REL2 signaling cassette in the expression of Vg, but not of Lp. We reveal molecular links that connect reproduction and immunity at several levels and provide a molecular basis for a long-suspected trade-off between these two processes.
2009
Blandin Stéphanie A, Wang-Sattler R, Lamacchia Marina, Gagneur Julien, Lycett G, Ning Y, Levashina Elena A, Steinmetz Lars M
Dissecting the genetic basis of resistance to malaria parasites in Anopheles gambiae Journal Article
In: Science, vol. 326, no. 5949, pp. 147-50, 2009.
Abstract | Links | BibTeX | Tags: blandin, M3i, TEP1
@article{SA2009,
title = {Dissecting the genetic basis of resistance to malaria parasites in Anopheles gambiae},
author = {Stéphanie A Blandin and R Wang-Sattler and Marina Lamacchia and Julien Gagneur and G Lycett and Y Ning and Elena A Levashina and Lars M Steinmetz},
url = {http://www.ncbi.nlm.nih.gov/pubmed/19797663},
year = {2009},
date = {2009-10-02},
journal = {Science},
volume = {326},
number = {5949},
pages = {147-50},
abstract = {The ability of Anopheles gambiae mosquitoes to transmit Plasmodium parasites is highly variable between individuals. However, the genetic basis of this variability has remained unknown. We combined genome-wide mapping and reciprocal allele-specific RNA interference (rasRNAi) to identify the genomic locus that confers resistance to malaria parasites and demonstrated that polymorphisms in a single gene encoding the antiparasitic thioester-containing protein 1 (TEP1) explain a substantial part of the variability in parasite killing. The link between TEP1 alleles and resistance to malaria may offer new tools for controlling malaria transmission. The successful application of rasRNAi in Anopheles suggests that it could also be applied to other organisms where RNAi is feasible to dissect complex phenotypes to the level of individual quantitative trait alleles.},
keywords = {blandin, M3i, TEP1},
pubstate = {published},
tppubtype = {article}
}
The ability of Anopheles gambiae mosquitoes to transmit Plasmodium parasites is highly variable between individuals. However, the genetic basis of this variability has remained unknown. We combined genome-wide mapping and reciprocal allele-specific RNA interference (rasRNAi) to identify the genomic locus that confers resistance to malaria parasites and demonstrated that polymorphisms in a single gene encoding the antiparasitic thioester-containing protein 1 (TEP1) explain a substantial part of the variability in parasite killing. The link between TEP1 alleles and resistance to malaria may offer new tools for controlling malaria transmission. The successful application of rasRNAi in Anopheles suggests that it could also be applied to other organisms where RNAi is feasible to dissect complex phenotypes to the level of individual quantitative trait alleles.
Fraiture Malou, Baxter Richard H G, Steinert Stefanie, Chelliah Yogarany, Frolet Cécile, Quispe-Tintaya Wilber, Hoffmann Jules A, Blandin Stéphanie A, Levashina Elena A
Two mosquito LRR proteins function as complement control factors in the TEP1-mediated killing of Plasmodium Journal Article
In: Cell Host Microbe, vol. 5, no. 3, pp. 273–284, 2009, ISSN: 1934-6069.
Abstract | Links | BibTeX | Tags: Animals, Anopheles, APL1, Biological, blandin, Complement System Proteins, Hemolymph, hoffmann, Immunologic Factors, LRIM1, M3i, Models, Plasmodium, Protein Binding, Proteins, TEP1
@article{fraiture_two_2009,
title = {Two mosquito LRR proteins function as complement control factors in the TEP1-mediated killing of Plasmodium},
author = {Malou Fraiture and Richard H G Baxter and Stefanie Steinert and Yogarany Chelliah and Cécile Frolet and Wilber Quispe-Tintaya and Jules A Hoffmann and Stéphanie A Blandin and Elena A Levashina},
doi = {10.1016/j.chom.2009.01.005},
issn = {1934-6069},
year = {2009},
date = {2009-03-01},
journal = {Cell Host Microbe},
volume = {5},
number = {3},
pages = {273--284},
abstract = {Plasmodium development within Anopheles mosquitoes is a vulnerable step in the parasite transmission cycle, and targeting this step represents a promising strategy for malaria control. The thioester-containing complement-like protein TEP1 and two leucine-rich repeat (LRR) proteins, LRIM1 and APL1, have been identified as major mosquito factors that regulate parasite loads. Here, we show that LRIM1 and APL1 are required for binding of TEP1 to parasites. RNAi silencing of the LRR-encoding genes results in deposition of TEP1 on Anopheles tissues, thereby depleting TEP1 from circulation in the hemolymph and impeding its binding to Plasmodium. LRIM1 and APL1 not only stabilize circulating TEP1, they also stabilize each other prior to their interaction with TEP1. Our results indicate that three major antiparasitic factors in mosquitoes jointly function as a complement-like system in parasite killing, and they reveal a role for LRR proteins as complement control factors.},
keywords = {Animals, Anopheles, APL1, Biological, blandin, Complement System Proteins, Hemolymph, hoffmann, Immunologic Factors, LRIM1, M3i, Models, Plasmodium, Protein Binding, Proteins, TEP1},
pubstate = {published},
tppubtype = {article}
}
Plasmodium development within Anopheles mosquitoes is a vulnerable step in the parasite transmission cycle, and targeting this step represents a promising strategy for malaria control. The thioester-containing complement-like protein TEP1 and two leucine-rich repeat (LRR) proteins, LRIM1 and APL1, have been identified as major mosquito factors that regulate parasite loads. Here, we show that LRIM1 and APL1 are required for binding of TEP1 to parasites. RNAi silencing of the LRR-encoding genes results in deposition of TEP1 on Anopheles tissues, thereby depleting TEP1 from circulation in the hemolymph and impeding its binding to Plasmodium. LRIM1 and APL1 not only stabilize circulating TEP1, they also stabilize each other prior to their interaction with TEP1. Our results indicate that three major antiparasitic factors in mosquitoes jointly function as a complement-like system in parasite killing, and they reveal a role for LRR proteins as complement control factors.
2008
Blandin Stéphanie A, Marois Eric, Levashina Elena A
Antimalarial responses in Anopheles gambiae: from a complement-like protein to a complement-like pathway Journal Article
In: Cell Host Microbe., vol. 3, no. 6, pp. 364-74, 2008.
Abstract | Links | BibTeX | Tags: blandin, M3i, marois, TEP1
@article{SA2008,
title = {Antimalarial responses in Anopheles gambiae: from a complement-like protein to a complement-like pathway},
author = {Stéphanie A Blandin and Eric Marois and Elena A Levashina},
url = {http://www.ncbi.nlm.nih.gov/pubmed/18541213},
year = {2008},
date = {2008-06-12},
journal = {Cell Host Microbe.},
volume = {3},
number = {6},
pages = {364-74},
abstract = {Malaria transmission between humans depends on the ability of Anopheles mosquitoes to support Plasmodium development. New perspectives in vector control are emerging from understanding the mosquito immune system, which plays critical roles in parasite recognition and killing. A number of factors controlling this process have been recently identified, and key among them is TEP1, a homolog of human complement factor C3 whose binding to the parasite surface targets it for subsequent killing. Here, we review our current knowledge of mosquito factors that respond to Plasmodium infection and elaborate on the activity and mode of action of the TEP1 complement-like pathway.},
keywords = {blandin, M3i, marois, TEP1},
pubstate = {published},
tppubtype = {article}
}
Malaria transmission between humans depends on the ability of Anopheles mosquitoes to support Plasmodium development. New perspectives in vector control are emerging from understanding the mosquito immune system, which plays critical roles in parasite recognition and killing. A number of factors controlling this process have been recently identified, and key among them is TEP1, a homolog of human