Elucidating mechanism cellular uptake removal

29 Jan

Viral vectors such as adenoviruses and retroviruses are commonly used in gene therapy owing to the high efficiency of cellular translocation as well as the non-selectivity toward cellular membrane [5,8].Although most of the viral vectors are uniquely suited for gene transfer as they can penetrate cell membranes via direct fusion or endocytosis, their routine use in both regular research laboratories and clinical settings are greatly limited due to safety concerns such as undesirable immune response, risk of tumorigenesis and insertional mutagenesis [8].The chemical structure of propylene glycol amine cationic lipid and PFBT are show in Figure S1.Lipofectamine 2000 was obtained from Invitrogen Ltd. Poly [(9,9-dioctylfluorenyl-2,7-diyl)-alt-co-(1,4-benzo-(2,1',3)-thiadiazole)] (PFBT) was purchased from American Dye Source, Inc. Dimethyl sulfoxide (DMSO), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), anhydrous tetrahydrofuran (THF) and other chemicals not mentioned were purchased from Sigma-Aldrich (St. The CPNPs were synthesized based on the nanoprecipitation method [29,30].State Key Laboratory of Medicinal Chemical Biology, Tianjin Key Laboratory of Biosensing and Molecular Recognition, College of Chemistry, Nankai University, Tianjin, 300071, China;3. with fluorescence quantum yield of 70.7±0.3%) and small size dimension of ~3.6±0.3 nm (DLS result).Hunan Provincial Key Laboratory of Food Science and Biotechnology, College of Food Science and Technology, Hunan Agricultural University, Changsha, 410128, China;4. Fast and efficient cellular translocation capability was observed according to the time-dependent living cell imaging experiments.PFBT) with zwitterionic lipid molecules (propylene glycol amine cationic lipid) together, positively charged CPNPs were obtained with ξ potential of 46.8 mv.

On this basis, the effectiveness of gene transfection is difficult to study without visualizing the exact transport noninvasively.As a consequence, exploring non-viral gene delivery materials has continuously received considerable attention because these materials can be structurally varied, are relatively safe, and have an ability to carry large and diverse genetic materials into cells.Until now, a number of studies involving non-viral gene delivery systems, such as cationic lipids having one or two hydrophobic tails, polycations, dendrimers and histones, were carried out [9-16].In comparison with the commonly used gene delivery vector, lipofectamine 2000 (with gene transfection efficiency of 55±5% for p EGFP), the gene expression efficiency with the positively charged CPNPs (70±3% for p EGFP) was improved significantly.Intracellular fluorescence imaging results demonstrated that the CPNPs could actively assemble close to the periphery of nuclei.