title: Folic acid conjugated mPEG-PEI600 as an efficient non-viral vector for targeted nucleic acid delivery.
abstract: In this study we describe a novel polymer, mPPS-FA, synthesized as a potential gene transfer vector. To complete mPPS-FA, folic acid was conjugated to a backbone (named mPPS) consisting of a copolymer of methyl PEG-2000, PEI-600, and sebacoyl chloride. (1)H NMR, FT-IR, and UV spectroscopy were used to characterize the structure of mPPS-FA. It was revealed that mPPS-FA holds the ability to bind plasmid DNA yielding positively charged particles (polyplexes). Dynamic light scattering (DLS) and TEM techniques were used to study the size and morphology of the formed mPPS-FA/DNA nanocomplexes. The mPPS-FA/DNA nanoparticles exhibited low cytotoxicity as transfection of B16-F0, U87MG, CHO-1, and Ho-8910 cells produced >80% viability indicating low cytotoxicity of the polymer. The ability of mPPS-FA to deliver EGFP plasmid to melanoma B16-F0, U87, CHO-1, Ho-8910, and A549 cells was investigated in vitro as compared to the lipid-based transfection agent Lipofectamine2000 and Linear PEI 22 kDa (L-PEI 22 kDa). We found that mPPS-FA/DNA complexes yielded the highest GFP transfection efficiency in B16-F0, U87, CHO-1, and Ho-8910 cells, which all highly express folate receptors (FR), at an mPPS-FA/DNA ratio (w/w) of 15. Furthermore, the transfection of mPPS-FA/DNA complexes in CHO-1 cells could be competitively blocked by free folic acid molecules. In contrast, in low FR expressing A549 cells, mPPS-FA showed similar low transfection efficiency as mPPS. Taken together, mPPS-FA showed the highest efficiency in vitro and the potential to be developed as a nonviral gene carrier.
authors.affiliation: Department of Chemistry, The University of Hong Kong, Pokfulam, Hong Kong, China; Brain Tumor Center, Neurosurgery, Faculty of Medicine, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong, China; School of Biomedical Science, The Chinese University of Hong Kong, Shatin, Hong Kong, China.
authors.forename: Zhenhua
authors.initials: Z
authors.lastname: Xu
authors.affiliation: Department of Chemistry, The University of Hong Kong, Pokfulam, Hong Kong, China.
authors.forename: Jiefu
authors.initials: J
authors.lastname: Jin
authors.affiliation: Department of Chemistry, The University of Hong Kong, Pokfulam, Hong Kong, China.
authors.forename: Leo K S
authors.initials: LKS
authors.lastname: Siu
authors.affiliation: Brain Tumor Center, Neurosurgery, Faculty of Medicine, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong, China; School of Biomedical Science, The Chinese University of Hong Kong, Shatin, Hong Kong, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Yao
authors.affiliation: Brain Tumor Center, Neurosurgery, Faculty of Medicine, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong, China.
authors.forename: Johnny
authors.initials: J
authors.lastname: Sze
authors.affiliation: Department of Chemistry, The University of Hong Kong, Pokfulam, Hong Kong, China.
authors.forename: Hongzhe
authors.initials: H
authors.lastname: Sun
authors.affiliation: School of Biomedical Science, The Chinese University of Hong Kong, Shatin, Hong Kong, China.
authors.forename: Hsiang-Fu
authors.initials: HF
authors.lastname: Kung
authors.affiliation: Brain Tumor Center, Neurosurgery, Faculty of Medicine, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong, China.
authors.forename: Wai Sang
authors.initials: WS
authors.lastname: Poon
authors.affiliation: Department of Chemistry, The University of Hong Kong, Pokfulam, Hong Kong, China; School of Biological Sciences, The University of Hong Kong, Hong Kong, China. Electronic address: ssmng@hku.hk.
authors.forename: Samuel S M
authors.initials: SSM
authors.lastname: Ng
authors.affiliation: Department of Chemistry, The University of Hong Kong, Pokfulam, Hong Kong, China; Brain Tumor Center, Neurosurgery, Faculty of Medicine, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong, China. Electronic address: mcllin@surgery.cuhk.edu.hk.
authors.forename: Marie C
authors.initials: MC
authors.lastname: Lin
mesh_terms: D000473:Alkanes / Q000737:chemistry* / Q000633:toxicity; D000818:Animals; D001667:Binding, Competitive; D016466:CHO Cells; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000737:chemistry / Q000378:metabolism*; D005492:Folic Acid / Q000737:chemistry / Q000378:metabolism* / Q000633:toxicity; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D007097:Imines / Q000737:chemistry* / Q000633:toxicity; D008027:Light; D009682:Magnetic Resonance Spectroscopy; D008745:Methylation; D051379:Mice; D046529:Microscopy, Electron, Transmission; D053758:Nanoparticles; D036103:Nanotechnology; D010316:Particle Size; D011092:Polyethylene Glycols / Q000737:chemistry* / Q000633:toxicity; D011095:Polyethylenes / Q000737:chemistry* / Q000633:toxicity; D012542:Scattering, Radiation; D013056:Spectrophotometry, Ultraviolet; D017550:Spectroscopy, Fourier Transform Infrared; D014162:Transfection / Q000379:methods*
chemical_list: D000473:Alkanes; D007097:Imines; D011095:Polyethylenes; C505405:poly(ethylene imine); C061659:sebacoyl chloride; D049452:Green Fluorescent Proteins; D011092:Polyethylene Glycols; D004247:DNA; D005492:Folic Acid