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首页> 外文期刊>The Journal of biological chemistry >Generation of a Drug-inducible Reporter System to Study Cell Reprogramming in Human Cells
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Generation of a Drug-inducible Reporter System to Study Cell Reprogramming in Human Cells

机译:生成一种药物诱导的报告系统,用于研究人体细胞的细胞重新编程

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Reprogramming of somatic cells into induced pluripotent stem cells is achieved by the expression of defined transcription factors. In the last few years, reprogramming strategies on the basis of doxycycline-inducible lentiviruses in mouse cells became highly powerful for screening purposes when the expression of a GFP gene, driven by the reactivation of endogenous stem cell specific promoters, was used as a reprogramming reporter signal. However, similar reporter systems in human cells have not been generated. Here, we describe the derivation of drug-inducible human fibroblast-like cell lines that express different subsets of reprogramming factors containing a GFP gene under the expression of the endogenous OCT4 promoter. These cell lines can be used to screen functional substitutes for reprogramming factors or modifiers of reprogramming efficiency. As a proof of principle of this system, we performed a screening of a library of pluripotent-enriched microRNAs and identified hsa-miR-519a as a novel inducer of reprogramming efficiency.
机译:通过表达定义的转录因子,实现了体细胞重编程到诱导多能干细胞中。在过去的几年中,在小鼠细胞中,在小鼠细胞的基础上重编程策略对于通过内源性干细胞特异性启动子的再激活驱动的GFP基因的表达时,在小鼠细胞中对小鼠细胞的表达具有高度强大的筛选目的。用作重编程报告者信号。然而,尚未产生人体细胞中的类似报道系统。在这里,我们描述了药物诱导的人成纤维细胞样细胞系的衍生,其在内源Oct4启动子的表达下表达含有GFP基因的重编程因子的不同子集。这些细胞系可用于筛选功能替代物用于重编程效率的重编程因子或改性剂。作为该系统原理的证据,我们进行了多能富含富含MicroRNA库的筛选,并将HSA-MIR-519A鉴定为重编程效率的新颖诱导剂。

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