Long-term efficient gene delivery using polyethylenimine with modified Tat peptide
Long-term efficient gene delivery using polyethylenimine with modified Tat peptide
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DOI：
10.1016/j.biomaterials.2013.11.012
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发表时间：
2014-02-01
期刊：
影响因子：
14
通讯作者：
Montclare, Jin K.
中科院分区：
文献类型：
--
作者：
Yamano, Seiichi;Dai, Jisen;Montclare, Jin K.
聚乙烯亚胺（PEI）是一种阳离子聚合物，作为一种高效的基因传递载体得到了广泛的研究和应用。同样，HIV-1 Tat肽，一种细胞渗透性肽，已经成功地用于细胞内基因传递。为了提高这两种载体的优良性能，我们将PEI与含有组氨酸和半胱氨酸残基（mTat）的修饰Tat肽序列结合起来。用荧光素酶表达质粒在两种细胞中评价mat / pei介导的体外转染。与单独使用mTat、单独使用PEI或四种商业试剂相比，mTat/PEI对两种细胞系的转染效率均有显著提高（约5倍），且细胞毒性很小。mTat/PEI/DNA复合物的粒径明显小于单独的mTat或PEI，这与更高的转染效率相关。Filipin III是一种小泡介导的内吞作用抑制剂，可显著抑制mTat/PEI转染。相比之下，氯丙嗪（一种网格蛋白介导的内吞作用抑制剂）则没有作用。这表明小泡介导的内吞作用是转染机制。此外，体内研究结果表明，肌内注射mTat/PEI/DNA的动物比单独注射mTat/DNA、PEI/DNA或DNA的动物荧光素酶表达明显更高、更长（7个月），且无任何相关毒性。mTat与PEI的结合可显著提高转染效率，扩大其作为非病毒基因载体在体内和体外的潜在应用。(C) 2013 Elsevier Ltd.版权所有。
Polyethylenimine (PEI), a cationic polymer, has been widely studied and shown great promise as an efficient gene delivery vehicle. Likewise, the HIV-1 Tat peptide, a cell-permeable peptide, has been successfully used for intracellular gene delivery. To improve the favorable properties of these two vectors, we combine PEI with the modified Tat peptide sequence bearing histidine and cysteine residues (mTat). In vitro mTat/PEI-mediated transfection was evaluated by luciferase expression plasmid in two cell types. mTat/PEI produced significant improvement (approximate to 5-fold) in transfection efficiency of both cell lines with little cytotoxicity when compared to mTat alone, PEI alone, or four commercial reagents. The particle size of mTat/PEI/DNA complex was significantly smaller than mTat or PEI alone, and it was correlated with higher transfection efficiency. Filipin III, an inhibitor of caveolae-mediated endocytosis, significantly inhibited mTat/PEI transfection. In contrast, chlorpromazine, an inhibitor of clathrin-mediated endocytosis, did not. This suggested caveolae-mediated endocytosis as the transfection mechanism. Furthermore, the results of in vivo studies showed that animals administered mTat/PEI/DNA intramuscularly had significantly higher and longer luciferase expression 7 months) than those with mTat/DNA, PEI/DNA, or DNA alone, without any associated toxicity. The combination of mTat with PEI could significantly improve transfection efficiency, expanding the potential use as a non-viral gene vector both in vitro and in vivo. (C) 2013 Elsevier Ltd. All rights reserved.