Reprogramming human fibroblasts using HIV-1 TAT recombinant proteins OCT4, SOX2, KLF4 and c-MYC.

Reprogramming human fibroblasts using HIV-1 TAT recombinant proteins OCT4, SOX2, KLF4 and c-MYC.
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使用 HIV-1 TAT 重组蛋白 OCT4、SOX2、KLF4 和 c-MYC 重编程人成纤维细胞

DOI:
10.1007/s11033-009-9680-6
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发表时间:
2010-04
影响因子:
2.8
通讯作者:
Bishop CE
Bishop CE
中科院分区:
生物学4区
文献类型:
--
作者:
Pan C;Lu B;Chen H;Bishop CE

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已经表明,人和鼠成纤维细胞可以通过使用病毒载体异位表达转录因子来重编程。出于人类治疗应用的目的,将病毒转基因整合到基因组中不太可能被接受。因此,我们在大肠杆菌(OCT4、SOX2、c-MYC 和 KLF4)中生产了携带 HIV1 细胞穿透 TAT 结构域的重组转录因子蛋白。当添加到组织培养基中时,纯化的蛋白质能够进入哺乳动物细胞,但似乎不会转位到细胞核。进一步的研究表明,大部分蛋白质被束缚在内体中,无法进行重编程。一旦这个问题得到解决,蛋白质重编程似乎将成为临床应用的首选方法。
It has been shown that human and murine fibroblasts can be reprogrammed by ectopic expression of transcription factors using viral vectors. For the purpose of human therapeutic applications, the integration of viral transgenes into the genome is unlikely to be accepted. We therefore produced recombinant transcription factor proteins in E. coli (OCT4, SOX2, c-MYC and KLF4) carrying the cell penetrating TAT domain from HIV1. The purified proteins were able to enter into mammalian cells when added to tissue culture medium but appeared not to translocate to the nucleus. Further investigation indicated that most of the protein was tied up in the endosomes and was unavailable for reprogramming. Once this problem has been solved it seems likely that protein reprogramming will be the method of choice for clinical applications.
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