title: Enhanced efficiency of P-element mediated transgenesis in Drosophila: Microinjection of DNA complexed with nanomaterial.
abstract: 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.
authors.affiliation: 1] CSIR-Indian Institute of Toxicology Research, Mahatma Gandhi Marg, Lucknow, UP, 226001, INDIA [2].
authors.forename: Madhavi
authors.initials: M
authors.lastname: Sonane
authors.forename: Ritu
authors.initials: R
authors.lastname: Goyal
authors.forename: Debapratim K
authors.initials: DK
authors.lastname: Chowdhuri
authors.forename: Kristipati Ravi
authors.initials: KR
authors.lastname: Ram
authors.forename: Kailash C
authors.initials: KC
authors.lastname: Gupta
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics; D004247:DNA / Q000008:administration & dosage* / Q000235:genetics*; D004330:Drosophila / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D008845:Microinjections / Q000379:methods; D053758:Nanoparticles / Q000008:administration & dosage; D049329:Nanostructures / Q000008:administration & dosage; D010957:Plasmids / Q000235:genetics; D011094:Polyethyleneimine / Q000008:administration & dosage; D014162:Transfection / Q000379:methods*; D019076:Transgenes / Q000235:genetics
chemical_list: D011094:Polyethyleneimine; D004247:DNA