人类多能干细胞的必需基因特征可鉴定出未知基因和底物依赖性。

Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies.

机构信息

Donnelly Centre, University of Toronto, Toronto, ON, Canada.

Donnelly Centre, University of Toronto, Toronto, ON, Canada; Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada.

出版信息

Cell Rep. 2019 Apr 9;27(2):599-615.e12. doi: 10.1016/j.celrep.2019.02.041.

DOI:10.1016/j.celrep.2019.02.041

PMID:30970261

Abstract

Human pluripotent stem cells (hPSCs) provide an invaluable tool for modeling diseases and hold promise for regenerative medicine. For understanding pluripotency and lineage differentiation mechanisms, a critical first step involves systematically cataloging essential genes (EGs) that are indispensable for hPSC fitness, defined as cell reproduction in this study. To map essential genetic determinants of hPSC fitness, we performed genome-scale loss-of-function screens in an inducible Cas9 H1 hPSC line cultured on feeder cells and laminin to identify EGs. Among these, we found FOXH1 and VENTX, genes that encode transcription factors previously implicated in stem cell biology, as well as an uncharacterized gene, C22orf43/DRICH1. hPSC EGs are substantially different from other human model cell lines, and EGs in hPSCs are highly context dependent with respect to different growth substrates. Our CRISPR screens establish parameters for genome-wide screens in hPSCs, which will facilitate the characterization of unappreciated genetic regulators of hPSC biology.

摘要

人类多能干细胞 (hPSCs) 为疾病建模提供了宝贵的工具，并且在再生医学方面有很大的应用潜力。为了了解多能性和谱系分化的机制，关键的第一步是系统地列出对 hPSC 适应性（在本研究中定义为细胞繁殖）必不可少的必需基因 (EGs)。为了绘制 hPSC 适应性的关键遗传决定因素图谱，我们在饲养细胞和层粘连蛋白上培养的诱导型 Cas9 H1 hPSC 系中进行了全基因组功能丧失筛选，以鉴定 EGs。在这些基因中，我们发现了 FOXH1 和 VENTX，它们编码转录因子，以前与干细胞生物学有关，以及一个未被描述的基因 C22orf43/DRICH1。hPSC 的 EGs 与其他人类模型细胞系有很大的不同，并且在不同的生长底物方面，hPSCs 的 EGs 具有高度的上下文依赖性。我们的 CRISPR 筛选为 hPSCs 的全基因组筛选建立了参数，这将有助于对 hPSC 生物学中未被认识的遗传调节剂进行表征。