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Enhanced efficiency of P-element mediated transgenesis in Drosophila: Microinjection of DNA complexed with nanomaterial

The efficiency of genetic transformation technology to generate stable transgenics depends upon the successful delivery of plasmid DNA in embryonic cells. The available gene vectors facilitate efficient plasmid DNA delivery to the cellular milieu but are exposed to nuclease degradation. Recent in vi...

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Autores principales: Sonane, Madhavi, Goyal, Ritu, Chowdhuri, Debapratim K., Ram, Kristipati Ravi, Gupta, Kailash C.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3847702/
https://www.ncbi.nlm.nih.gov/pubmed/24296454
http://dx.doi.org/10.1038/srep03408
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author Sonane, Madhavi
Goyal, Ritu
Chowdhuri, Debapratim K.
Ram, Kristipati Ravi
Gupta, Kailash C.
author_facet Sonane, Madhavi
Goyal, Ritu
Chowdhuri, Debapratim K.
Ram, Kristipati Ravi
Gupta, Kailash C.
author_sort Sonane, Madhavi
collection PubMed
description The efficiency of genetic transformation technology to generate stable transgenics depends upon the successful delivery of plasmid DNA in embryonic cells. The available gene vectors facilitate efficient plasmid DNA delivery to the cellular milieu but are exposed to nuclease degradation. Recent in vitro studies suggest encapsulation of plasmid DNA with nanomaterial(s) for better protection against nucleases. Therefore, in this study, we tested if complexing of free plasmid DNA with linear polyethylenimine (LPEI, 25 kDa) based nanoparticle (LPN) enhances the efficiency of transformation (transgenesis) by using Drosophila based germ-line transformation technology. Here, we show that the LPN-DNA complex not only enhances the efficiency of this transgenic technology at a DNA concentration of 0.04 μg/μl but also reduces the DNA quantity required to generate transgenics by ten folds. This approach has potential applications for other types of transgenesis and nucleic acid injection methods in Drosophila as well as other popular genetic model systems.
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spelling pubmed-38477022013-12-03 Enhanced efficiency of P-element mediated transgenesis in Drosophila: Microinjection of DNA complexed with nanomaterial Sonane, Madhavi Goyal, Ritu Chowdhuri, Debapratim K. Ram, Kristipati Ravi Gupta, Kailash C. Sci Rep Article The efficiency of genetic transformation technology to generate stable transgenics depends upon the successful delivery of plasmid DNA in embryonic cells. The available gene vectors facilitate efficient plasmid DNA delivery to the cellular milieu but are exposed to nuclease degradation. Recent in vitro studies suggest encapsulation of plasmid DNA with nanomaterial(s) for better protection against nucleases. Therefore, in this study, we tested if complexing of free plasmid DNA with linear polyethylenimine (LPEI, 25 kDa) based nanoparticle (LPN) enhances the efficiency of transformation (transgenesis) by using Drosophila based germ-line transformation technology. Here, we show that the LPN-DNA complex not only enhances the efficiency of this transgenic technology at a DNA concentration of 0.04 μg/μl but also reduces the DNA quantity required to generate transgenics by ten folds. This approach has potential applications for other types of transgenesis and nucleic acid injection methods in Drosophila as well as other popular genetic model systems. Nature Publishing Group 2013-12-03 /pmc/articles/PMC3847702/ /pubmed/24296454 http://dx.doi.org/10.1038/srep03408 Text en Copyright © 2013, Macmillan Publishers Limited. All rights reserved http://creativecommons.org/licenses/by-nc-sa/3.0/ This work is licensed under a Creative Commons Attribution-NonCommercial-ShareALike 3.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-sa/3.0/
spellingShingle Article
Sonane, Madhavi
Goyal, Ritu
Chowdhuri, Debapratim K.
Ram, Kristipati Ravi
Gupta, Kailash C.
Enhanced efficiency of P-element mediated transgenesis in Drosophila: Microinjection of DNA complexed with nanomaterial
title Enhanced efficiency of P-element mediated transgenesis in Drosophila: Microinjection of DNA complexed with nanomaterial
title_full Enhanced efficiency of P-element mediated transgenesis in Drosophila: Microinjection of DNA complexed with nanomaterial
title_fullStr Enhanced efficiency of P-element mediated transgenesis in Drosophila: Microinjection of DNA complexed with nanomaterial
title_full_unstemmed Enhanced efficiency of P-element mediated transgenesis in Drosophila: Microinjection of DNA complexed with nanomaterial
title_short Enhanced efficiency of P-element mediated transgenesis in Drosophila: Microinjection of DNA complexed with nanomaterial
title_sort enhanced efficiency of p-element mediated transgenesis in drosophila: microinjection of dna complexed with nanomaterial
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3847702/
https://www.ncbi.nlm.nih.gov/pubmed/24296454
http://dx.doi.org/10.1038/srep03408
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