title: Selective blocking of primary amines in branched polyethylenimine with biocompatible ligand alleviates cytotoxicity and augments gene delivery efficacy in mammalian cells.
abstract: Recently, polyethylenimines (PEIs) have emerged as efficient vectors for nucleic acids delivery. However, inherent cytotoxicity has limited their in vivo applications. To address this concern as well as to incorporate hydrophobic domains for improving interactions with the lipid bilayers in the cell membranes, we have tethered varying amounts of amphiphilic pyridoxyl moieties onto bPEI to generate a small series of pyridoxyl-PEI (PyP) polymers. Spectroscopic characterization confirms the formation of PyP polymers, which subsequently form stable complexes with pDNA in nanometric range with positive surface charge. The projected modification not only accounts for a decrease in the density of 1° amines but also allows formation of relatively loose complexes with pDNA (cf. bPEI). Alleviation of the cytotoxicity, efficient interaction with cell membranes and easy disassembly of the pDNA complexes have led to the remarkable enhancement in the transfection efficiency of PyP/pDNA complexes in mammalian cells with one of the formulations, PyP-3/pDNA complex, showing transfection in ∼68% cells compared to ∼16% cells by Lipofectamine/pDNA complex. Further, the efficacy of PyP-3 vector has been established by delivering GFP-specific siRNA resulting in ∼88% suppression of the target gene expression. These results demonstrate the efficacy of the projected carriers that can be used in future gene therapy applications.
authors: Nucleic Acids Research Laboratory, CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, Delhi 110007, India.
authors: Sushil K
authors: SK
authors: Tripathi
authors: Nucleic Acids Research Laboratory, CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, Delhi 110007, India.
authors: Niharika
authors: N
authors: Gupta
authors: Nucleic Acids Research Laboratory, CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, Delhi 110007, India.
authors: Manohar
authors: M
authors: Mahato
authors: Nucleic Acids Research Laboratory, CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, Delhi 110007, India; CSIR-Indian Institute of Toxicology Research, M.G. Marg, Lucknow 226001, India.
authors: Kailash C
authors: KC
authors: Gupta
authors: Nucleic Acids Research Laboratory, CSIR-Institute of Genomics and Integrative Biology, Delhi University Campus, Mall Road, Delhi 110007, India. Electronic address: pkumar@igib.res.in.
authors: Pradeep
authors: P
authors: Kumar
mesh_terms: D000588:Amines / Q000737:chemistry*; D001672:Biocompatible Materials / Q000494:pharmacology*; D002021:Buffers; D016923:Cell Death / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000378:metabolism; D005453:Fluorescence; D018014:Gene Transfer Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006367:HeLa Cells; D006461:Hemolysis / Q000187:drug effects; D006493:Heparin / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008055:Lipids / Q000737:chemistry; D061986:MCF-7 Cells; D023282:Nuclease Protection Assays; D010316:Particle Size; D010957:Plasmids / Q000378:metabolism; D011094:Polyethyleneimine / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D034741:RNA, Small Interfering / Q000378:metabolism; D055672:Static Electricity; D017186:Titrimetry
chemical_list: D000588:Amines; D001672:Biocompatible Materials; D002021:Buffers; D008024:Ligands; D008055:Lipids; C086724:Lipofectamine; D034741:RNA, Small Interfering; D049452:Green Fluorescent Proteins; D011094:Polyethyleneimine; D006493:Heparin; D004247:DNA; D003850:Deoxyribonuclease I
keywords: Buffering capacity; Cytotoxicity; Polyethylenimine; Pyridoxal hydrochloride; Transfection efficiency