http://scholars.ntou.edu.tw/handle/123456789/8622
標題: | Dependence of transgene expression and the relative buffering capacity of dextran-grafted polyethylenimine | 作者: | Tseng, Wen-Chi Fang, Tsuei-Yun Su, Ling-Yu Tang, Chien-Hsiang |
關鍵字: | transfection;delivery;conjugation;nonviral vector;cellular mechanism | 公開日期: | 五月-2005 | 卷: | 2 | 期: | 3 | 來源出版物: | Molecular Pharmaceutics | 摘要: | Branched polyethylenimine (PEI) is a cationic polymer capable of forming self-assembly complexes with DNA to become a highly efficient agent used in gene delivery. Conjugation through the primary amines of PEI is a most commonly used approach further to enable the targeting delivery or to improve the stability of the DNA-polymer complexes. An understanding of how the conjugation affects the transfection mechanisms can help in the design of efficient polycationic vectors. In order to investigate the effects of conjugation, folate and the dextrans of molecular weight 1500 (dex-1500) and 10 000 (dex-10000) were used to prepare three different types of PEI conjugates: dextran-PEI, folate-PEI, and folate-dextran-PEI, which were subsequently employed to form complexes with DNA. These conjugates were found to cause less cytotoxicity than the unmodified PEI as revealed by the MTT method, and to be able to deliver an approximate amount of ethidium monoazide labeled plasmid into the cells. The efficiencies of green fluorescent protein (GFP) expression mediated by these conjugates, however, were less efficient than those mediated by the unmodified PEI. A titration experiment suggested that conjugation through the primary amines of PEI resulted in the loss of relative buffering capacity, a major factor aiding the release of plasmid from the endosomes, presumably because the conjugated molecules hindered the protonation of the PEI conjugates. When a quantitative relationship between relative buffering capacity and transfection efficiency was examined, a threshold of relative buffering capacity, around 50% of the unmodified PEI, was noted to be required for minimal detection of GFP positive cells. In addition, the cytotoxicity could be also related to the relative buffering capacity in an approximately linear trend. It is thus concluded that the severe loss of relative buffering capacity by conjugation might be attributed to the inefficiency of transgene expression mediated by the dextran-PEI conjugates. |
URI: | http://scholars.ntou.edu.tw/handle/123456789/8622 | ISSN: | 1543-8384 | DOI: | 10.1021/mp050007t |
顯示於: | 食品科學系 |
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