Quantitative aspects of endocytic activity in lipid-mediated transfections.
Variation in transfection efficiency observed in different cell-types is poorly understood. To investigate the influence of endocytic activity on lipid-mediated transfections, we have monitored both the processes in 12 different cell-types. The endocytic activity shows a strong positive correlation (P < 0.01), with transfection efficiency. Treatment with wortmannin resulted in cell-type-dependent inhibition of transfection. Studies on M-phase cells by confocal microscopy show that compared to interphase cells, uptake of cationic liposomes was substantially reduced. In addition, transfection efficiency of cells in mitotic phase was inhibited by >70% compared to controls. Our study based on several cell-types demonstrates for the first time that quantitative aspects of endocytosis have decisive influence on the overall process of lipid-mediated transgene expression.
Prasad
Tekkatte Krishnamurthy
TK
Centre for Cellular and Molecular Biology, Hyderabad, India. tkprasad1975@gmail.com
Rangaraj
Nandini
N
Rao
Nalam Madhusudhana
NM
D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D006690:Bisbenzimidazole; D016466:CHO Cells; D019556:COS Cells; D002455:Cell Division; D002460:Cell Line; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D006224:Cricetinae; D003412:Cricetulus; D004705:Endocytosis / Q000187:drug effects* / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005229:Fatty Acids, Monounsaturated / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D017930:Genes, Reporter; D006367:HeLa Cells; D006801:Humans; D007739:L Cells; D008081:Liposomes; D051379:Mice; D018613:Microscopy, Confocal; D016328:NF-kappa B / Q000032:analysis / Q000378:metabolism; D041681:NIH 3T3 Cells; D010714:Phosphatidylethanolamines / Q000378:metabolism; D000081082:Phosphoinositide-3 Kinase Inhibitors; D000644:Quaternary Ammonium Compounds / Q000378:metabolism; D012235:Rhodamines; D014162:Transfection; D000077191:Wortmannin
C410262:1,2-dioleoyl-glycero-3-phosphatidyl ethanolamine; D000730:Androstadienes; D004791:Enzyme Inhibitors; D005229:Fatty Acids, Monounsaturated; D005456:Fluorescent Dyes; D008081:Liposomes; D016328:NF-kappa B; D010714:Phosphatidylethanolamines; D000081082:Phosphoinositide-3 Kinase Inhibitors; D000644:Quaternary Ammonium Compounds; D012235:Rhodamines; D016650:Fluorescein-5-isothiocyanate; D006690:Bisbenzimidazole; C070046:1,2-dioleoyloxy-3-(trimethylammonium)propane; D000077191:Wortmannin