title: Efficient targeted pDNA/siRNA delivery with folate-low-molecular-weight polyethyleneimine-modified pullulan as non-viral carrier.
abstract: Folate receptor (FR)-mediated gene/short interfering RNA (siRNA) targeting shows advantage for the delivery of gene/siRNA into specific FR-overexpressing cancer cells. In this study, the non-targeted gene vector P-PEI was synthesized by grafting low-molecular-weight (1kDa) branched polyethyleneimine (PEI) to succinylated pullulan, and the targeted gene vector P-PEI-FA was synthesized by coupling the carboxyl of folate (FA) to the amino of PEI. Gel electrophoresis retardation assay demonstrated that both P-PEI and P-PEI-FA can efficiently wrap pDNA and siRNA with electrostatic interaction at N/P ratios higher than 1.56 and can protect pDNA from degradation by DNase I and serum. Compared with PEI/pDNA, P-PEI/pDNA and P-PEI-FA/pDNA showed lower cytotoxicity against different cells. Under serum-containing conditions, compared with Lipofamine 2000/DNA and Lipofamine2000/siRNA, P-PEI-FA/DNA at N/P ratio of 6.25 displayed higher gene transfection efficiency, whereas P-PEI-FA/siRNA at N/P ratio of 12.5 demonstrated better enhanced gene silencing effect. P-PEI-FA/siRNA can also deliver FAM-labeled siRNA to endosomes and escape. Moreover, the gene transfection and silencing effects of P-PEI-FA were higher than those of P-PEI, and were dependent on the dose of FA in FR(+) HeLa cells. Thus, P-PEI-FA can assist DNA or siRNA targeting to FR-overexpressing cells, and the uptake pathway of P-PEI-FA/siRNA was FR-mediated endocytosis. These results indicate that P-PEI-FA is a potential candidate for safe and targeted gene delivery applications.
authors.affiliation: State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024, PR China; School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, PR China. Electronic address: wangjingyun67@dlut.edu.cn.
authors.forename: Jingyun
authors.initials: J
authors.lastname: Wang
authors.forename: Bairui
authors.initials: B
authors.lastname: Dou
authors.forename: Yongming
authors.initials: Y
authors.lastname: Bao
mesh_terms: D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000378:metabolism*; D003850:Deoxyribonuclease I / Q000378:metabolism; D004337:Drug Carriers; D004587:Electrophoresis, Agar Gel; D005434:Flow Cytometry; D005492:Folic Acid / Q000494:pharmacology*; D020868:Gene Silencing / Q000187:drug effects; D018014:Gene Transfer Techniques; D005936:Glucans / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006801:Humans; D042541:Intracellular Space / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D023282:Nuclease Protection Assays; D010957:Plasmids / Q000378:metabolism*; D011094:Polyethyleneimine / Q000138:chemical synthesis / Q000737:chemistry*; D034741:RNA, Small Interfering / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet; D014162:Transfection
chemical_list: D004337:Drug Carriers; D005936:Glucans; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; C009109:pullulan; D011094:Polyethyleneimine; D004247:DNA; D005492:Folic Acid; D003850:Deoxyribonuclease I
keywords: Folate; Non-virus gene vector; Polyethyleneimine; Pullulan; siRNA