title: Polyglutamic acid-based nanocomposites as efficient non-viral gene carriers in vitro and in vivo.
abstract: A series of polyethylenimine (PEI) and γ-polyglutamic acid (PGA) nanocomposites (PPGA) was prepared and evaluated in terms of their cell viability and transfection efficiency in vitro and in vivo. On complexion with pDNA, the positively charged PPGA/DNA nanocomposites resulted in a higher level of in vitro reporter gene transfection (2.7-7.9-fold) as compared to native PEI, and selected commercial reagents and >95% cell viability in HEK293, HeLa and HepG2 cell lines. Further, PPGA-5 nanocomposite (the best working system in terms of transfection efficiency among the series) was found to efficiently transfect primary mouse keratinocytes up to 22% above the control level. PPGA-5, when tested for in vivo cytotoxicity in Drosophila, did not induce any stress in the exposed larvae in comparison with control. In vivo gene expression using PPGA-5 showed the highest transfection efficiency in spleen of mouse closely followed by heart tissues after intravenous injection through tail vein. Besides, these nanocomposites also delivered siRNA efficiently into mammalian cells, resulting in ∼ 80% suppression of EGFP expression. These results together demonstrated the potential of the projected nanocomposites for in vivo gene delivery.
authors.affiliation: CSIR - Institute of Genomics and Integrative Biology, Delhi University Campus, Delhi, India.
authors.forename: Sushil Kumar
authors.initials: SK
authors.lastname: Tripathi
authors.forename: R
authors.initials: R
authors.lastname: Goyal
authors.forename: K M
authors.initials: KM
authors.lastname: Ansari
authors.forename: K
authors.initials: K
authors.lastname: Ravi Ram
authors.forename: Y
authors.initials: Y
authors.lastname: Shukla
authors.forename: D K
authors.initials: DK
authors.lastname: Chowdhuri
authors.forename: K C
authors.initials: KC
authors.lastname: Gupta
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000008:administration & dosage / Q000235:genetics; D004330:Drosophila / Q000187:drug effects; D004337:Drug Carriers / Q000737:chemistry* / Q000633:toxicity; D005260:Female; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism; D049409:Luciferases, Firefly / Q000235:genetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D053761:Nanocomposites; D010957:Plasmids; D011094:Polyethyleneimine / Q000737:chemistry / Q000633:toxicity; D011099:Polyglutamic Acid / Q000737:chemistry* / Q000633:toxicity; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics; D013154:Spleen / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D055672:Static Electricity; D014162:Transfection
chemical_list: D004337:Drug Carriers; D034741:RNA, Small Interfering; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D011099:Polyglutamic Acid; D011094:Polyethyleneimine; D004247:DNA; D049409:Luciferases, Firefly