Genome-wide CRISPR-KO Screen Uncovers mTORC1-Mediated Gsk3 Regulation in Naive Pluripotency Maintenance and Dissolution

Cell Rep. 2018 Jul 10;24(2):489-502. doi: 10.1016/j.celrep.2018.06.027.

Abstract

The genetic basis of naive pluripotency maintenance and loss is a central question in embryonic stem cell biology. Here, we deploy CRISPR-knockout-based screens in mouse embryonic stem cells to interrogate this question through a genome-wide, non-biased approach using the Rex1GFP reporter as a phenotypic readout. This highly sensitive and efficient method identified genes in diverse biological processes and pathways. We uncovered a key role for negative regulators of mTORC1 in maintenance and exit from naive pluripotency and provided an integrated account of how mTORC1 activity influences naive pluripotency through Gsk3. Our study therefore reinforces Gsk3 as the central node and provides a comprehensive, data-rich resource that will improve our understanding of mechanisms regulating pluripotency and stimulate avenues for further mechanistic studies.

Keywords: Akt; CRISPR; GATOR1; Nprl2; Tsc2; exit from pluripotency; mTORC1; mTORC2; naive pluripotency; screening.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / genetics
  • Cell Self Renewal
  • Clustered Regularly Interspaced Short Palindromic Repeats / genetics*
  • Gene Expression Regulation, Developmental
  • Gene Knockout Techniques*
  • Genome*
  • Glycogen Synthase Kinase 3 / metabolism
  • Mechanistic Target of Rapamycin Complex 1 / metabolism*
  • Mice
  • Mice, Knockout
  • Models, Biological
  • Mouse Embryonic Stem Cells
  • Phenotype
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism*
  • Transcriptome / genetics

Substances

  • Mechanistic Target of Rapamycin Complex 1
  • Glycogen Synthase Kinase 3