Mapping Germ-layer Specification Preventing Genes in HPSCs Via Genome-scale CRISPR Screening

Overview

Journal iScience

Publisher Cell Press

Date 2021 Jan 1

PMID 33385119

Citations 5

Authors

Xiangjie Xu

Yanhua Du

Lin Ma

Shuwei Zhang

Lei Shi

Zhenyu Chen

Zhongshu Zhou

Yi Hui

Yang Liu

Yujiang Fang

Beibei Fan

Zhongliang Liu

Nan Li

Shanshan Zhou

Cizhong Jiang

Ling Liu

Xiaoqing Zhang

Affiliations

Soon will be listed here.

Abstract

Understanding the biological processes that determine the entry of three germ layers of human pluripotent stem cells (hPSCs) is a central question in developmental and stem cell biology. Here, we genetically engineered hPSCs with the germ layer reporter and inducible CRISPR/Cas9 knockout system, and a genome-scale screening was performed to define pathways restricting germ layer specification. Genes clustered in the key biological processes, including embryonic development, mRNA processing, metabolism, and epigenetic regulation, were centered in the governance of pluripotency and lineage development. Other than typical pluripotent transcription factors and signaling molecules, loss of function of mesendodermal specifiers resulted in advanced neuroectodermal differentiation, given their inter-germ layer antagonizing effect. Regarding the epigenetic superfamily, microRNAs enriched in hPSCs showed clear germ layer-targeting specificity. The cholesterol synthesis pathway maintained hPSCs via retardation of neuroectoderm specification. Thus, in this study, we identified a full landscape of genetic wiring and biological processes that control hPSC self-renewal and trilineage specification.

Citing Articles

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