title: Gellan gum blended PEI nanocomposites as gene delivery agents: evidences from in vitro and in vivo studies.
abstract: Branched Polyethylenimine, 25 kDa (PEI), was blended with gellan gum, an anionic heteropolysaccharide, for partial neutralization of its excess positive charge to form gellan gum-polyethylenimine (GP) nanocomposites (NCs). Subsequently, we manipulated the amount of gellan gum for obtaining a series of NCs and characterized them for their size, charge and morphology. Among all the NCs, one member, named GP3, showed the best transfection efficiency in tested cell lines in comparison with the rest of the series, PEI, Lipofectamine and other commercial transfection agents and also exhibited minimum cytotoxicity. It was found to transfect primary cells of mouse skin with better efficiency than PEI and Lipofectamine and was able to protect the plasmid DNA from nucleases and serum proteins present in the blood. GP3 exhibited efficient intracellular delivery of plasmid as revealed by confocal studies while its intracellular presence was also confirmed by the knockdown of GFP expression (using GFP specific siRNA) and JNKII by quantifying proteins in cell lysates and by western blotting and hybridization, respectively. In vivo cytotoxicity studies in Drosophila showed lack of induction of stress response in the exposed organisms. Further, exposed organisms did not show any developmental delay or mortality and no morphological defects were observed in the emerged flies. In vivo gene expression studies in Balb/c mice revealed maximum expression of luciferase enzyme in spleen. The study suggests that GP3 may act as an efficient non-viral gene carrier with diverse biomedical applications.
authors.affiliation: Institute of Genomics and Integrative Biology (CSIR), Delhi University Campus, Delhi, India.
authors.forename: Ritu
authors.initials: R
authors.lastname: Goyal
authors.forename: S K
authors.initials: SK
authors.lastname: Tripathi
authors.forename: Shilpa
authors.initials: S
authors.lastname: Tyagi
authors.forename: K
authors.initials: K
authors.lastname: Ravi Ram
authors.forename: K M
authors.initials: KM
authors.lastname: Ansari
authors.forename: Y
authors.initials: Y
authors.lastname: Shukla
authors.forename: D
authors.initials: D
authors.lastname: Kar Chowdhuri
authors.forename: Pradeep
authors.initials: P
authors.lastname: Kumar
authors.forename: K C
authors.initials: KC
authors.lastname: Gupta
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D004247:DNA / Q000032:analysis / Q000378:metabolism; D016503:Drug Delivery Systems; D004353:Drug Evaluation, Preclinical; D005079:Excipients / Q000737:chemistry; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D056945:Hep G2 Cells; D006801:Humans; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D053761:Nanocomposites / Q000032:analysis / Q000737:chemistry*; D010957:Plasmids; D011094:Polyethyleneimine / Q000737:chemistry* / Q000378:metabolism; D011135:Polysaccharides, Bacterial / Q000737:chemistry* / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection; D023041:Xenograft Model Antitumor Assays
chemical_list: D005079:Excipients; D011135:Polysaccharides, Bacterial; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; C048288:gellan gum; D011094:Polyethyleneimine; D004247:DNA