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Generation of a Drug-inducible Reporter System to Study Cell Reprogramming in Human Cells

机译:药物诱导报告系统的生成用于研究人细胞中的细胞重编程

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摘要

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:通过定义的转录因子的表达来实现将体细胞重编程为诱导的多能干细胞。在过去的几年中,当将由内源性干细胞特异性启动子重新激活驱动的GFP基因的表达用作重编程报告基因时,基于强力霉素可诱导的慢病毒在小鼠细胞中的重编程策略变得非常强大,可用于筛选信号。但是,尚未产生人类细胞中类似的报告系统。在这里,我们描述了药物诱导的人类成纤维细胞样细胞系的衍生,该细胞系在内源OCT4启动子的表达下表达包含GFP基因的重编程因子的不同子集。这些细胞系可用于筛选重编程因子的功能替代物或重编程效率的调节剂。作为该系统原理的证明,我们筛选了富含多能性microRNA的文库,并将hsa-miR-519a鉴定为重编程效率的新型诱导剂。

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