Mapping the Effects of Genetic Variation on Chromatin State and Gene Expression Reveals Loci That Control Ground State Pluripotency

Cell Stem Cell. 2020 Sep 3;27(3):459-469.e8. doi: 10.1016/j.stem.2020.07.005. Epub 2020 Aug 13.

Abstract

Mouse embryonic stem cells (mESCs) cultured in the presence of LIF occupy a ground state with highly active pluripotency-associated transcriptional and epigenetic circuitry. However, ground state pluripotency in some inbred strain backgrounds is unstable in the absence of ERK1/2 and GSK3 inhibition. Using an unbiased genetic approach, we dissect the basis of this divergent response to extracellular cues by profiling gene expression and chromatin accessibility in 170 genetically heterogeneous mESCs. We map thousands of loci affecting chromatin accessibility and/or transcript abundance, including 10 QTL hotspots where genetic variation at a single locus coordinates the regulation of genes throughout the genome. For one hotspot, we identify a single enhancer variant ∼10 kb upstream of Lifr associated with chromatin accessibility and mediating a cascade of molecular events affecting pluripotency. We validate causation through reciprocal allele swaps, demonstrating the functional consequences of noncoding variation in gene regulatory networks that stabilize pluripotent states in vitro.

Keywords: Diversity Outbred; Lifr; chromatin accessibility; gene expression; genetic variation; ground state pluripotency; mESC; metastability; quantitative trait locus.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation
  • Chromatin* / genetics
  • Gene Expression
  • Genetic Variation
  • Glycogen Synthase Kinase 3
  • Mice
  • Pluripotent Stem Cells*

Substances

  • Chromatin
  • Glycogen Synthase Kinase 3