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2023
de Faria Isaque J. S., Imler Jean-Luc, Marques João T.
Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies Article de journal
Dans: STAR Protocols, vol. 4, iss. 1, 2023.
Résumé | Liens | BibTeX | Étiquettes: antibody, cell bioloby, Immunology, M3i, Marques, microbiology, Microscopy, Molecular Biology
@article{deFaria2023,
title = {Protocol for the analysis of double-stranded RNAs in virus-infected insect cells using anti-dsRNA antibodies},
author = {Isaque J.S. de Faria and Jean-Luc Imler and João T. Marques},
url = {https://doi.org/10.1016/j.xpro.2022.102033},
doi = {10.1016/j.xpro.2022.102033},
year = {2023},
date = {2023-03-17},
urldate = {2023-03-17},
journal = {STAR Protocols},
volume = {4},
issue = {1},
abstract = {Characterization of double-stranded (ds)RNAs is relevant to the understanding of viral replication and immune sensing. Here, we provide a protocol describing the use of anti-dsRNA antibodies for immunofluorescence and immunoblotting in virus-infected insect cells, which can also be applied to tissues and other organisms. We describe the procedures to prepare insect cells for viral infection, followed by RNA extraction and in vitro production of synthetic dsRNA controls. We then detail the steps for dsRNA detection by immunoblotting and immunofluorescence. For complete details on the use and execution of this protocol, please refer to de Faria et al. (2022).1},
keywords = {antibody, cell bioloby, Immunology, M3i, Marques, microbiology, Microscopy, Molecular Biology},
pubstate = {published},
tppubtype = {article}
}
Characterization of double-stranded (ds)RNAs is relevant to the understanding of viral replication and immune sensing. Here, we provide a protocol describing the use of anti-dsRNA antibodies for immunofluorescence and immunoblotting in virus-infected insect cells, which can also be applied to tissues and other organisms. We describe the procedures to prepare insect cells for viral infection, followed by RNA extraction and in vitro production of synthetic dsRNA controls. We then detail the steps for dsRNA detection by immunoblotting and immunofluorescence. For complete details on the use and execution of this protocol, please refer to de Faria et al. (2022).1
2022
de Faria Isaque J. S., Aguiar Eric R. G. R., Olmo Roenick P., da Silva Juliana Alves, Daeffler Laurent, Carthew Richard W., Imler Jean-Luc, Marques Joao T.
Invading viral DNA triggers dsRNA synthesis by RNA polymerase II to activate antiviral RNA interference in Drosophila Article de journal
Dans: Cell Reports, vol. 39, p. 110976, 2022.
Résumé | Liens | BibTeX | Étiquettes: antiviral, Drosophila, dsRNA, imler, M3i, Marques, protocol, RNA Interference
@article{dedaMarques2022,
title = {Invading viral DNA triggers dsRNA synthesis by RNA polymerase II to activate antiviral RNA interference in Drosophila},
author = {Isaque J.S. de Faria and Eric R.G.R. Aguiar and Roenick P. Olmo and Juliana Alves da Silva and Laurent Daeffler and Richard W. Carthew and Jean-Luc Imler and Joao T. Marques},
doi = {10.1016/j.celrep.2022.110976},
year = {2022},
date = {2022-06-21},
urldate = {2022-06-21},
journal = {Cell Reports},
volume = {39},
pages = {110976},
abstract = {dsRNA sensing triggers antiviral responses against RNA and DNA viruses in diverse eukaryotes. In Drosophila, Invertebrate iridescent virus 6 (IIV-6), a large DNA virus, triggers production of small interfering RNAs (siRNAs) by the dsRNA sensor Dicer-2. Here, we show that host RNA polymerase II (RNAPII) bidirec- tionally transcribes specific AT-rich regions of the IIV-6 DNA genome to generate dsRNA. Both replicative and naked IIV-6 genomes trigger production of dsRNA in Drosophila cells, implying direct sensing of invading DNA. Loquacious-PD, a Dicer-2 co-factor essential for the biogenesis of endogenous siRNAs, is dispensable for processing of IIV-6-derived dsRNAs, which suggests that they are distinct. Consistent with this finding, inhibition of the RNAPII co-factor P-TEFb affects the synthesis of endogenous, but not virus-derived, dsRNA. Altogether, our results suggest that a non-canonical RNAPII complex recognizes invading viral DNA to synthesize virus-derived dsRNA, which activates the antiviral siRNA pathway in Drosophila.},
keywords = {antiviral, Drosophila, dsRNA, imler, M3i, Marques, protocol, RNA Interference},
pubstate = {published},
tppubtype = {article}
}
dsRNA sensing triggers antiviral responses against RNA and DNA viruses in diverse eukaryotes. In Drosophila, Invertebrate iridescent virus 6 (IIV-6), a large DNA virus, triggers production of small interfering RNAs (siRNAs) by the dsRNA sensor Dicer-2. Here, we show that host RNA polymerase II (RNAPII) bidirec- tionally transcribes specific AT-rich regions of the IIV-6 DNA genome to generate dsRNA. Both replicative and naked IIV-6 genomes trigger production of dsRNA in Drosophila cells, implying direct sensing of invading DNA. Loquacious-PD, a Dicer-2 co-factor essential for the biogenesis of endogenous siRNAs, is dispensable for processing of IIV-6-derived dsRNAs, which suggests that they are distinct. Consistent with this finding, inhibition of the RNAPII co-factor P-TEFb affects the synthesis of endogenous, but not virus-derived, dsRNA. Altogether, our results suggest that a non-canonical RNAPII complex recognizes invading viral DNA to synthesize virus-derived dsRNA, which activates the antiviral siRNA pathway in Drosophila.
2021
Leite Thiago H J F, Ferreira Alvaro G A, Imler Jean-Luc, Marques João T
Distinct Roles of Hemocytes at Different Stages of Infection by Dengue and Zika Viruses in Aedes aegypti Mosquitoes Article de journal
Dans: Frontiers in immunology, vol. 12, p. 660873, 2021.
Résumé | Liens | BibTeX | Étiquettes: Aedes, Dengue, Hemocytes, imler, innate immunity, M3i, Marques, Zika
@article{Leite2021,
title = {Distinct Roles of Hemocytes at Different Stages of Infection by Dengue and Zika Viruses in Aedes aegypti Mosquitoes},
author = {Thiago H J F Leite and Alvaro G A Ferreira and Jean-Luc Imler and João T Marques},
url = {https://www.frontiersin.org/articles/10.3389/fimmu.2021.660873/full},
doi = {10.3389/fimmu.2021.660873},
year = {2021},
date = {2021-05-13},
journal = {Frontiers in immunology},
volume = {12},
pages = {660873},
abstract = {Aedes aegypti mosquitoes are vectors for arboviruses of medical importance such as dengue (DENV) and Zika (ZIKV) viruses. Different innate immune pathways contribute to the control of arboviruses in the mosquito vector including RNA interference, Toll and Jak- STAT pathways. However, the role of cellular responses mediated by circulating macrophage-like cells known as hemocytes remains unclear. Here we show that hemocytes are recruited to the midgut of Ae. aegypti mosquitoes in response to DENV or ZIKV. Blockade of the phagocytic function of hemocytes using latex beads induced increased accumulation of hemocytes in the midgut and a reduction in virus infection levels in this organ. In contrast, inhibition of phagocytosis by hemocytes led to increased systemic dissemination and replication of DENV and ZIKV. Hence, our work reveals a dual role for hemocytes in Ae. aegypti mosquitoes, whereby phagocytosis is not required to control viral infection in the midgut but is essential to restrict systemic dissemination. Further understanding of the mechanism behind this duality could help the design of vector-based strategies to prevent transmission of arboviruses.},
keywords = {Aedes, Dengue, Hemocytes, imler, innate immunity, M3i, Marques, Zika},
pubstate = {published},
tppubtype = {article}
}
Aedes aegypti mosquitoes are vectors for arboviruses of medical importance such as dengue (DENV) and Zika (ZIKV) viruses. Different innate immune pathways contribute to the control of arboviruses in the mosquito vector including RNA interference, Toll and Jak- STAT pathways. However, the role of cellular responses mediated by circulating macrophage-like cells known as hemocytes remains unclear. Here we show that hemocytes are recruited to the midgut of Ae. aegypti mosquitoes in response to DENV or ZIKV. Blockade of the phagocytic function of hemocytes using latex beads induced increased accumulation of hemocytes in the midgut and a reduction in virus infection levels in this organ. In contrast, inhibition of phagocytosis by hemocytes led to increased systemic dissemination and replication of DENV and ZIKV. Hence, our work reveals a dual role for hemocytes in Ae. aegypti mosquitoes, whereby phagocytosis is not required to control viral infection in the midgut but is essential to restrict systemic dissemination. Further understanding of the mechanism behind this duality could help the design of vector-based strategies to prevent transmission of arboviruses.
2020
Cai H, Holleufer A, Simonsen B, Schneider J, Lemoine A, Gad HH, Huang J, Huang J, Chen D, Peng T, Marques JT, Hartmann R, Martins N, Imler JL
2'3'-cGAMP triggers a STING- and NF-κB-dependent broad antiviral response in Drosophila Article de journal
Dans: Science Signaling, vol. 13, no. 660, p. eabc4537, 2020.
Résumé | Liens | BibTeX | Étiquettes: imler, M3i, Marques, STING
@article{Cai_2020,
title = {2'3'-cGAMP triggers a STING- and NF-κB-dependent broad antiviral response in Drosophila},
author = {H Cai and A Holleufer and B Simonsen and J Schneider and A Lemoine and HH Gad and J Huang and J Huang and D Chen and T Peng and JT Marques and R Hartmann and N Martins and JL Imler},
url = {https://pubmed.ncbi.nlm.nih.gov/33262294/},
doi = {10.1126/scisignal.abc4537 },
year = {2020},
date = {2020-12-01},
journal = {Science Signaling},
volume = {13},
number = {660},
pages = {eabc4537},
abstract = {We previously reported that an ortholog of STING regulates infection by picorna-like viruses in Drosophila In mammals, STING is activated by the cyclic dinucleotide 2'3'-cGAMP produced by cGAS, which acts as a receptor for cytosolic DNA. Here, we showed that injection of flies with 2'3'-cGAMP induced the expression of dSTING-regulated genes. Coinjection of 2'3'-cGAMP with a panel of RNA or DNA viruses resulted in substantially reduced viral replication. This 2'3'-cGAMP-mediated protection was still observed in flies with mutations in Atg7 and AGO2, genes that encode key components of the autophagy and small interfering RNA pathways, respectively. By contrast, this protection was abrogated in flies with mutations in the gene encoding the NF-κB transcription factor Relish. Transcriptomic analysis of 2'3'-cGAMP-injected flies revealed a complex response pattern in which genes were rapidly induced, induced after a delay, or induced in a sustained manner. Our results reveal that dSTING regulates an NF-κB-dependent antiviral program that predates the emergence of interferons in vertebrates. },
keywords = {imler, M3i, Marques, STING},
pubstate = {published},
tppubtype = {article}
}
We previously reported that an ortholog of STING regulates infection by picorna-like viruses in Drosophila In mammals, STING is activated by the cyclic dinucleotide 2'3'-cGAMP produced by cGAS, which acts as a receptor for cytosolic DNA. Here, we showed that injection of flies with 2'3'-cGAMP induced the expression of dSTING-regulated genes. Coinjection of 2'3'-cGAMP with a panel of RNA or DNA viruses resulted in substantially reduced viral replication. This 2'3'-cGAMP-mediated protection was still observed in flies with mutations in Atg7 and AGO2, genes that encode key components of the autophagy and small interfering RNA pathways, respectively. By contrast, this protection was abrogated in flies with mutations in the gene encoding the NF-κB transcription factor Relish. Transcriptomic analysis of 2'3'-cGAMP-injected flies revealed a complex response pattern in which genes were rapidly induced, induced after a delay, or induced in a sustained manner. Our results reveal that dSTING regulates an NF-κB-dependent antiviral program that predates the emergence of interferons in vertebrates.
Aguiar ERGR, de Almeida JPP, Queiroz LR, Oliveira LS, Olmo RP, de Faria IJDS, Imler JL, Gruber A, Matthews BJ, Marques JT
A single unidirectional piRNA cluster similar to the flamenco locus is the major source of EVE-derived transcription and small RNAs in Aedes aegypti mosquitoes Article de journal
Dans: RNA, vol. 26, no. 5, p. 581-594, 2020.
Résumé | Liens | BibTeX | Étiquettes: A. aegypti, Aedes aegypti, endogenous viral elements, EVE, flamenco locus, imler, M3i, Marques, piRNA, piRNAs, RNA Interference
@article{Aguiar_2020,
title = {A single unidirectional piRNA cluster similar to the flamenco locus is the major source of EVE-derived transcription and small RNAs in Aedes aegypti mosquitoes },
author = {ERGR Aguiar and JPP de Almeida and LR Queiroz and LS Oliveira and RP Olmo and IJDS de Faria and JL Imler and A Gruber and BJ Matthews and JT Marques},
url = {https://rnajournal.cshlp.org/content/26/5/581.long},
doi = {10.1261/rna.073965.119},
year = {2020},
date = {2020-01-29},
journal = {RNA},
volume = {26},
number = {5},
pages = {581-594},
abstract = {Endogenous viral elements (EVEs) are found in many eukaryotic genomes. Despite considerable knowledge about genomic elements such as transposons (TEs) and retroviruses, we still lack information about nonretroviral EVEs. Aedes aegypti mosquitoes have a highly repetitive genome that is covered with EVEs. Here, we identified 129 nonretroviral EVEs in the AaegL5 version of the A. aegypti genome. These EVEs were significantly associated with TEs and preferentially located in repeat-rich clusters within intergenic regions. Genome-wide transcriptome analysis showed that most EVEs generated transcripts although only around 1.4% were sense RNAs. The majority of EVE transcription was antisense and correlated with the generation of EVE-derived small RNAs. A single genomic cluster of EVEs located in a 143 kb repetitive region in chromosome 2 contributed with 42% of antisense transcription and 45% of small RNAs derived from viral elements. This region was enriched for TE-EVE hybrids organized in the same coding strand. These generated a single long antisense transcript that correlated with the generation of phased primary PIWI-interacting RNAs (piRNAs). The putative promoter of this region had a conserved binding site for the transcription factor Cubitus interruptus, a key regulator of the flamenco locus in Drosophila melanogaster Here, we have identified a single unidirectional piRNA cluster in the A. aegypti genome that is the major source of EVE transcription fueling the generation of antisense small RNAs in mosquitoes. We propose that this region is a flamenco-like locus in A. aegypti due to its relatedness to the major unidirectional piRNA cluster in Drosophila melanogaster. },
keywords = {A. aegypti, Aedes aegypti, endogenous viral elements, EVE, flamenco locus, imler, M3i, Marques, piRNA, piRNAs, RNA Interference},
pubstate = {published},
tppubtype = {article}
}
Endogenous viral elements (EVEs) are found in many eukaryotic genomes. Despite considerable knowledge about genomic elements such as transposons (TEs) and retroviruses, we still lack information about nonretroviral EVEs. Aedes aegypti mosquitoes have a highly repetitive genome that is covered with EVEs. Here, we identified 129 nonretroviral EVEs in the AaegL5 version of the A. aegypti genome. These EVEs were significantly associated with TEs and preferentially located in repeat-rich clusters within intergenic regions. Genome-wide transcriptome analysis showed that most EVEs generated transcripts although only around 1.4% were sense RNAs. The majority of EVE transcription was antisense and correlated with the generation of EVE-derived small RNAs. A single genomic cluster of EVEs located in a 143 kb repetitive region in chromosome 2 contributed with 42% of antisense transcription and 45% of small RNAs derived from viral elements. This region was enriched for TE-EVE hybrids organized in the same coding strand. These generated a single long antisense transcript that correlated with the generation of phased primary PIWI-interacting RNAs (piRNAs). The putative promoter of this region had a conserved binding site for the transcription factor Cubitus interruptus, a key regulator of the flamenco locus in Drosophila melanogaster Here, we have identified a single unidirectional piRNA cluster in the A. aegypti genome that is the major source of EVE transcription fueling the generation of antisense small RNAs in mosquitoes. We propose that this region is a flamenco-like locus in A. aegypti due to its relatedness to the major unidirectional piRNA cluster in Drosophila melanogaster.
2019
Olmo RP, Martins NE, Aguiar ERGR, Marques JT, Imler JL
The insect reservoir of biodiversity for viruses and for antiviral mechanisms Article de journal
Dans: An Acad Bras Cienc , vol. 91, no. Suppl 3, p. e20190122, 2019.
Résumé | Liens | BibTeX | Étiquettes: antiviral immunity, imler, insects, M3i, Marques, metagenomics, restriction factors, RNA Interference, virome
@article{Olmo_2019,
title = {The insect reservoir of biodiversity for viruses and for antiviral mechanisms},
author = {RP Olmo and NE Martins and ERGR Aguiar and JT Marques and JL Imler},
url = {https://www.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37652019000600604&lng=en&nrm=iso&tlng=en},
doi = {10.1590/0001-3765201920190122},
year = {2019},
date = {2019-06-03},
journal = { An Acad Bras Cienc },
volume = {91},
number = {Suppl 3},
pages = {e20190122},
abstract = {Insects are the most diverse group of animals. They can be infected by an extraordinary diversity of viruses. Among them, arthropod-borne viruses (arboviruses) can be transmitted to humans. High-throughput sequencing of small RNAs from insects provides insight into their virome, which may help understand the dynamics of vector borne infectious diseases. Furthermore, investigating the mechanisms that restrict viral infections in insects points to genetic innovations that may inspire novel antiviral strategies. },
keywords = {antiviral immunity, imler, insects, M3i, Marques, metagenomics, restriction factors, RNA Interference, virome},
pubstate = {published},
tppubtype = {article}
}
Insects are the most diverse group of animals. They can be infected by an extraordinary diversity of viruses. Among them, arthropod-borne viruses (arboviruses) can be transmitted to humans. High-throughput sequencing of small RNAs from insects provides insight into their virome, which may help understand the dynamics of vector borne infectious diseases. Furthermore, investigating the mechanisms that restrict viral infections in insects points to genetic innovations that may inspire novel antiviral strategies.
Martins NE, Olmo RP, Aguiar ERGR, Marques JT, Imler JL
Les insectes : un fantastique réservoir de virus et de gènes antiviraux Article de journal
Dans: Biologie Aujourd'hui, vol. 212, no. 3-4, p. 101-106, 2019.
Résumé | Liens | BibTeX | Étiquettes: antiviral immunity, ARN interference, imler, Insect, M3i, Marques, metagenomic, virome
@article{Martins_2019jbio,
title = {Les insectes : un fantastique réservoir de virus et de gènes antiviraux},
author = {NE Martins and RP Olmo and ERGR Aguiar and JT Marques and JL Imler},
url = {https://www.biologie-journal.org/articles/jbio/abs/2018/02/jbio190008/jbio190008.html},
doi = {10.1051/jbio/2019008},
year = {2019},
date = {2019-04-11},
journal = {Biologie Aujourd'hui},
volume = {212},
number = {3-4},
pages = {101-106},
abstract = {Les insectes forment le groupe d’animaux qui présente la plus grande diversité. Des travaux récents de métagénomique montrent qu’ils peuvent être infectés par une diversité extraordinaire de virus. Parmi eux, les arbovirus (arthropod-borne viruses) peuvent être transmis à l’Homme par les insectes hématophages, notamment les moustiques. Le séquençage à haut débit des petits ARN des insectes fournit des informations sur leur virome, un paramètre qui pourrait contribuer à expliquer la dynamique de la transmission des maladies infectieuses par des insectes vecteurs. D’autre part, la caractérisation des mécanismes qui restreignent les infections virales chez les insectes révèle des innovations génétiques qui pourraient à terme inspirer de nouvelles stratégies antivirales.},
keywords = {antiviral immunity, ARN interference, imler, Insect, M3i, Marques, metagenomic, virome},
pubstate = {published},
tppubtype = {article}
}
Les insectes forment le groupe d’animaux qui présente la plus grande diversité. Des travaux récents de métagénomique montrent qu’ils peuvent être infectés par une diversité extraordinaire de virus. Parmi eux, les arbovirus (arthropod-borne viruses) peuvent être transmis à l’Homme par les insectes hématophages, notamment les moustiques. Le séquençage à haut débit des petits ARN des insectes fournit des informations sur leur virome, un paramètre qui pourrait contribuer à expliquer la dynamique de la transmission des maladies infectieuses par des insectes vecteurs. D’autre part, la caractérisation des mécanismes qui restreignent les infections virales chez les insectes révèle des innovations génétiques qui pourraient à terme inspirer de nouvelles stratégies antivirales.
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 Article de journal
Dans: Nature Microbiology, vol. 3, no. 12, p. 1385-1393, 2018.
Résumé | Liens | BibTeX | Étiquettes: 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.
