I2CT

@article{singh_binding_2005,

title = {Binding and condensation of plasmid DNA onto functionalized carbon nanotubes: toward the construction of nanotube-based gene delivery vectors},

author = {Ravi Singh and Davide Pantarotto and David McCarthy and Olivier Chaloin and Johan Hoebeke and Charalambos D Partidos and Jean-Paul Briand and Maurizio Prato and Alberto Bianco and Kostas Kostarelos},

doi = {10.1021/ja0441561},

issn = {0002-7863},

year  = {2005},

date = {2005-03-01},

journal = {Journal of the American Chemical Society},

volume = {127},

number = {12},

pages = {4388--4396},

abstract = {Carbon nanotubes (CNTs) constitute a class of nanomaterials that possess characteristics suitable for a variety of possible applications. Their compatibility with aqueous environments has been made possible by the chemical functionalization of their surface, allowing for exploration of their interactions with biological components including mammalian cells. Functionalized CNTs (f-CNTs) are being intensively explored in advanced biotechnological applications ranging from molecular biosensors to cellular growth substrates. We have been exploring the potential of f-CNTs as delivery vehicles of biologically active molecules in view of possible biomedical applications, including vaccination and gene delivery. Recently we reported the capability of ammonium-functionalized single-walled CNTs to penetrate human and murine cells and facilitate the delivery of plasmid DNA leading to expression of marker genes. To optimize f-CNTs as gene delivery vehicles, it is essential to characterize their interactions with DNA. In the present report, we study the interactions of three types of f-CNTs, ammonium-functionalized single-walled and multiwalled carbon nanotubes (SWNT-NH3+; MWNT-NH3+), and lysine-functionalized single-walled carbon nanotubes (SWNT-Lys-NH3+), with plasmid DNA. Nanotube-DNA complexes were analyzed by scanning electron microscopy, surface plasmon resonance, PicoGreen dye exclusion, and agarose gel shift assay. The results indicate that all three types of cationic carbon nanotubes are able to condense DNA to varying degrees, indicating that both nanotube surface area and charge density are critical parameters that determine the interaction and electrostatic complex formation between f-CNTs with DNA. All three different f-CNT types in this study exhibited upregulation of marker gene expression over naked DNA using a mammalian (human) cell line. Differences in the levels of gene expression were correlated with the structural and biophysical data obtained for the f-CNT:DNA complexes to suggest that large surface area leading to very efficient DNA condensation is not necessary for effective gene transfer. However, it will require further investigation to determine whether the degree of binding and tight association between DNA and nanotubes is a desirable trait to increase gene expression efficiency in vitro or in vivo. This study constitutes the first thorough investigation into the physicochemical interactions between cationic functionalized carbon nanotubes and DNA toward construction of carbon nanotube-based gene transfer vector systems.},

keywords = {carbon, Cations, DNA, Electron, Gene Transfer Techniques, Genetic Vectors, I2CT, Lysine, Microscopy, Nanotubes, Plasmids, Quaternary Ammonium Compounds, scanning, Surface Plasmon Resonance, Team-Bianco},

pubstate = {published},

tppubtype = {article}

}

@article{de_marco_solubility_2004,

title = {The solubility and stability of recombinant proteins are increased by their fusion to NusA},

author = {Valeria De Marco and Gunter Stier and Stephanie A Blandin and Ario de Marco},

doi = {10.1016/j.bbrc.2004.07.189},

issn = {0006-291X},

year  = {2004},

date = {2004-01-01},

journal = {Biochem. Biophys. Res. Commun.},

volume = {322},

number = {3},

pages = {766--771},

abstract = {The new bacterial vector pETM60 enables the expression of His-tagged recombinant proteins fused to the C-terminus of NusA through a TEV protease recognition sequence. Three sequences coding for two protein domains (Xklp3A and Tep3Ag) and one membrane-bound viral protein (E8R) could not be expressed in a soluble form in bacteria. Their GST-fusions were mostly soluble but quickly degraded during purification. The same sequences cloned in pETM60 were efficiently purified by metal affinity and recovered soluble after the removal of the fusion partner. The NusA-fused constructs enabled to yield 13-20mg of fusion protein per litre of culture and 2.5-5mg of pure protein per litre of culture. Structural analysis indicated that the purified proteins were monodispersed and correctly folded. NusA has been used to raise antibodies that have been successfully used for Western blot and immunoprecipitation of NusA fusion proteins.},

keywords = {blandin, Drug Stability, Escherichia coli Proteins, Genetic Vectors, Glutathione Transferase, Kinetics, M3i, Oxidation-Reduction, Peptide Elongation Factors, Recombinant Fusion Proteins, Recombinant Proteins, Solubility, Transcription Factors, Transcriptional Elongation Factors},

pubstate = {published},

tppubtype = {article}

}

@article{reichhart_splice-activated_2002,

title = {Splice-activated UAS hairpin vector gives complete RNAi knockout of single or double target transcripts in Drosophila melanogaster},

author = {Jean-Marc Reichhart and Petros Ligoxygakis and Silvia Naitza and Gertrud Woerfel and Jean-Luc Imler and David Gubb},

doi = {10.1002/gene.10122},

issn = {1526-954X},

year  = {2002},

date = {2002-01-01},

journal = {Genesis (New York, N.Y.: 2000)},

volume = {34},

number = {1-2},

pages = {160--164},

keywords = {Animals, DNA Transposable Elements, DNA-Binding Proteins, Enhancer Elements, Genetic, Genetic Vectors, Genetically Modified, imler, M3i, reichhart, Saccharomyces cerevisiae Proteins, Transcription Factors},

pubstate = {published},

tppubtype = {article}

}