Development of poly(amino ester glycol urethane)/siRNA polyplexes for gene silencing.
Nonviral vectors, with their low immunogenicity and lack of pathogenicity, offer significant promise for siRNA therapy with fewer safety concerns. Nonviral vectors were also preferred in most transient siRNA delivery due to their ease of preparation. Previously, we incorporated tertiary amines and polyethylene glycol (PEG) into poly(ester urethane) to synthesize a soluble poly(amino ester glycol urethane), PaE(G)U, as a novel DNA transfection reagent for transgene delivery. The aim of this study was to develop PaE(G)U/siRNA polyplexes for gene silencing. We characterized the properties of PaE(G)U/siRNA polyplexes and compared them with those of PaE(G)U/DNA polyplexes. Using the Alexa Fluor 488-labeled, nonsilencing control siRNA as the reporter, we visualized cellular uptake of PaE(G)U/siRNA polyplexes and optimized the mass ratio of PaE(G)U/siRNA for delivery at 80/1. At this ratio, the average diameter of polyplexes was 540 nm, which was significantly larger than the average diameter of PaE(G)U/DNA polyplexes at 155 nm for efficient DNA delivery. Using the optimized PaE(G)U/siRNA polyplexes, transient silencing of constitutive luciferase expression (up to 92%) was achieved in our recombinant human HT-1080 fibroblast model via anti-luciferase siRNA delivery. In conclusion, PaE(G)U/siRNA polyplexes were developed and optimized for cellular uptake to allow efficient gene silencing. Engineering of soluble biodegradable polymers to incorporate amino, ester, PEG, and urethane units in the backbone constitutes a useful approach for the future design of siRNA carriers.
Tseng
S-Ja
SJ
Department of Chemical Engineering, National Tsing Hua University, 101, Sec. 2, Kuang Fu Road, Hsinchu 300, Taiwan.
Tang
Shiue-Cheng
SC
D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D001672:Biocompatible Materials / Q000138:chemical synthesis* / Q000494:pharmacology; D004337:Drug Carriers / Q000138:chemical synthesis* / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism / Q000473:pathology; D020868:Gene Silencing; D018014:Gene Transfer Techniques; D006801:Humans; D007700:Kinetics; D008156:Luciferases / Q000276:immunology / Q000378:metabolism; D008954:Models, Biological; D011140:Polyurethanes / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000737:chemistry*; D014162:Transfection / Q000379:methods*
D000906:Antibodies; D001672:Biocompatible Materials; D004337:Drug Carriers; D011140:Polyurethanes; D034741:RNA, Small Interfering; C521588:poly(amino ester glycol urethane); D008156:Luciferases