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Gene Editing to Generate Versatile Human Pluripotent Stem Cell Reporter Lines for Analysis of Differentiation and Lineage Tracing

机译:基因编辑以生成用于分化分析和谱系追踪的多功能人类多能干细胞报告基因系

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摘要

Abstract Transcription factors (TFs) are potent proteins that control gene expression and can thereby drive cell fate decisions. Fluorescent reporters have been broadly knocked into endogenous TF loci to investigate the biological roles of these factors; however, the sensitivity of such analyses in human pluripotent stem cells (hPSCs) is often compromised by low TF expression levels and/or reporter silencing. Complementarily, we report an inducible and quantitative reporter platform based on the Cre‐LoxP recombination system that enables robust, quantifiable, and continuous monitoring of live hPSCs and their progeny to investigate the roles of TFs during human development and disease. Stem Cells 2019;37:1556–1566
机译:摘要 转录因子(TFs)是控制基因表达的有效蛋白质,可以驱动细胞命运决策。荧光报告基因已被广泛地敲入内源性TF基因座,以研究这些因子的生物学作用;然而,此类分析在人类多能干细胞 (hPSC) 中的灵敏度通常受到低 TF 表达水平和/或报告基因沉默的影响。作为补充,我们报告了一种基于Cre-LoxP重组系统的诱导型定量报告基因平台,该平台能够对活的hPSC及其后代进行稳健、可量化和连续的监测,以研究TF在人类发育和疾病中的作用。干细胞 2019;37:1556–1566

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