Cell transfection in vitro and in vivo with nontoxic TAT peptide-liposome-DNA complexes
Cell transfection in vitro and in vivo with nontoxic TAT peptide-liposome-DNA complexes
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DOI：
10.1073/pnas.0435906100
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发表时间：
2003-02-18
影响因子：
11.1
通讯作者：
D'Souza, GGM
中科院分区：
文献类型：
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作者：
Torchilin, VP;Levchenko, TS;D'Souza, GGM
用TAT多肽修饰的脂质体(TATP-脂质体)显示出快速有效的移位到细胞质，随后迁移到核周区。含有少量(小于或等于10摩尔%)阳离子脂的TATP-脂质体与DNA形成牢固的非共价复合体。在这里，我们展示了TATP-脂质体-DNA复合体在体外和体内的转染结果。体外将该复合体导入小鼠NIH/3T3成纤维细胞和大鼠H9C2心肌细胞。TATP-脂质体相关的绿色荧光蛋白(GFP)基因载体pEGFP-N1的转染率高，但细胞毒性低于常用的阳离子脂质体基因传递系统。瘤内注射TATP-脂质体-DNA复合体也能诱导肿瘤细胞表达GFP。该系统可用于各种细胞治疗方案的体外和体外以及体内的局部基因治疗。
Liposomes modified with TAT peptide (TATp-liposomes) showed fast and efficient translocation into the cell cytoplasm with subsequent migration into the perinuclear zone. TATp-liposomes containing a small quantity (less than or equal to10 mol %) of a cationic lipid formed firm noncovalent complexes with DNA. Here, we present results demonstrating both in vitro and in vivo transfection with TATp-liposome-DNA complexes. Mouse NIH/3T3 fibroblasts and rat H9C2 cardiomyocytes were transfected with such complexes in vitro. The transfection with the TATp-liposome-associated pEGFP-N1 plasmid encoding for the green fluorescent protein (GFP) was high, whereas the cytotoxicity was lower than that of commonly used cationic lipid-based gene-delivery systems. Intratumoral injection of TATp-liposome-DNA complexes into the Lewis lung carcinoma tumor of mice also resulted in an expression of GFP in tumor cells. This transfection system should be useful for various protocols of cell treatment in vitro or ex vivo as well as for localized in vivo gene therapy.