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基因编辑生成多功能人多能干细胞报告系,用于分化分析和谱系示踪。

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

机构信息

Department of Bioengineering, University of California, Berkeley, California, USA.

Department of Chemical and Biomolecular Engineering, University of California, Berkeley, California, USA.

出版信息

Stem Cells. 2019 Dec;37(12):1556-1566. doi: 10.1002/stem.3096. Epub 2019 Nov 13.

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.

摘要

转录因子(TFs)是一种能够控制基因表达的强效蛋白,从而可以驱动细胞命运决定。荧光报告基因已广泛敲入内源性 TF 基因座,以研究这些因子的生物学作用;然而,在人多能干细胞(hPSCs)中,此类分析的灵敏度通常因 TF 表达水平低和/或报告基因沉默而受到影响。作为补充,我们报告了一种基于 Cre-LoxP 重组系统的诱导型和定量报告基因平台,该平台能够对 hPSCs 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。Stem Cells 2019;37:1556-1566.

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