Molecular weight-dependent genetic information transfer with disulfide-linked polyethylenimine-based nonviral vectors.
One strategy for improving gene vector properties of polyethylenimine is to facilitate individual transfection mechanism steps. This study investigates (i) improving transfection efficiency by attaching peptide nuclear localization signals (nuclear localization signals: SV40 large T antigen nuclear localization signal or C-terminus of histone H1) to polyethylenimine (10 kDa) and (ii) using disulfide linkages, which are expected to be stable during polyplex formation, but cleaved inside cells giving improved gene release. Nuclear localization signal-containing polyplexes exhibited low cytotoxicity, whereas transfection efficiency with high molecular weight plasmid DNA increased up to 3.6 times that of underivatized polyethylenimine in Neuro2A cells at higher molar ratio of polyethylenimine-nitrogen to DNA-phosphate (N/P) ratios. However, with luciferase-specific low molecular weight small interfering RNA in Neuro2A/EGFPLuc cells, nuclear localization signal-containing polyplexes with disulfide linkages caused substantial cytotoxicity at N/P ratios >15 and no consistent significant reduction in luciferase expression. Possible explanations for molecular weight-dependent differences in genetic information transfer by polyplexes containing disulfide-linked nuclear localization signals are discussed.
Pharmaceutical Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
Hamideh
H
Parhiz
Maryam
M
Hashemi
Arash
A
Hatefi
Wayne Thomas
WT
Shier
Sara Amel
SA
Farzad
Mohammad
M
Ramezani
D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry / Q000633:toxicity; D002460:Cell Line; D004220:Disulfides / Q000737:chemistry; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000737:chemistry* / Q000633:toxicity; D008156:Luciferases / Q000235:genetics; D008422:Materials Testing; D051379:Mice; D008970:Molecular Weight; D019913:Nuclear Localization Signals / Q000235:genetics; D011094:Polyethyleneimine / Q000737:chemistry* / Q000633:toxicity; D034741:RNA, Small Interfering / Q000235:genetics; D014162:Transfection
D001672:Biocompatible Materials; D004220:Disulfides; D019913:Nuclear Localization Signals; D034741:RNA, Small Interfering; D011094:Polyethyleneimine; D008156:Luciferases