Ash2l interacts with Oct4-stemness circuitry to promote super-enhancer-driven pluripotency network

Nucleic Acids Res. 2019 Nov 4;47(19):10115-10133. doi: 10.1093/nar/gkz801.


Pluripotency and cell fates can be modulated through the regulation of super-enhancers; however, the underlying mechanisms are unclear. Here, we showed a novel mechanism in which Ash2l directly binds to super-enhancers of several stemness genes to regulate pluripotency and self-renewal in pluripotent stem cells. Ash2l recruits Oct4/Sox2/Nanog (OSN) to form Ash2l/OSN complex at the super-enhancers of Jarid2, Nanog, Sox2 and Oct4, and further drives enhancer activation, upregulation of stemness genes, and maintains the pluripotent circuitry. Ash2l knockdown abrogates the OSN recruitment to all super-enhancers and further hinders the enhancer activation. In addition, CRISPRi/dCas9-mediated blocking of Ash2l-binding motifs at these super-enhancers also prevents OSN recruitment and enhancer activation, validating that Ash2l directly binds to super-enhancers and initiates the pluripotency network. Transfection of Ash2l with W118A mutation to disrupt Ash2l-Oct4 interaction fails to rescue Ash2l-driven enhancer activation and pluripotent gene upregulation in Ash2l-depleted pluripotent stem cells. Together, our data demonstrated Ash2l formed an enhancer-bound Ash2l/OSN complex that can drive enhancer activation, govern pluripotency network and stemness circuitry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • CRISPR-Cas Systems / genetics
  • Cell Differentiation / genetics
  • Cell Lineage / genetics
  • Cell Self Renewal / genetics
  • Cellular Reprogramming / genetics
  • DNA-Binding Proteins / genetics*
  • Enhancer Elements, Genetic* / genetics
  • Gene Expression Regulation, Developmental / genetics
  • Humans
  • Mice
  • Mouse Embryonic Stem Cells / metabolism*
  • Mutation / genetics
  • Nanog Homeobox Protein / genetics
  • Octamer Transcription Factor-3 / genetics*
  • Pluripotent Stem Cells / metabolism
  • SOXB1 Transcription Factors / genetics
  • Transcription Factors / genetics*
  • Transfection


  • Ash2l protein, mouse
  • DNA-Binding Proteins
  • Nanog Homeobox Protein
  • Nanog protein, mouse
  • Octamer Transcription Factor-3
  • Pou5f1 protein, mouse
  • SOXB1 Transcription Factors
  • Sox2 protein, mouse
  • Transcription Factors