Down-regulation of ATF1 leads to early neuroectoderm differentiation of human embryonic stem cells by increasing the expression level of SOX2

FASEB J. 2019 Sep;33(9):10577-10592. doi: 10.1096/fj.201800220RR. Epub 2019 Jun 26.

Abstract

We reveal by high-throughput screening that activating transcription factor 1 (ATF1) is a novel pluripotent regulator in human embryonic stem cells (hESCs). The knockdown of ATF1 expression significantly up-regulated neuroectoderm (NE) genes but not mesoderm, endoderm, and trophectoderm genes. Of note, down-regulation or knockout of ATF1 with short hairpin RNA (shRNA), small interfering RNA (siRNA), or clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) was sufficient to up-regulate sex-determining region Y-box (SOX)2 and paired box 6 (PAX6) expression under the undifferentiated or differentiated conditions, whereas overexpression of ATF1 suppressed NE differentiation. Endogenous ATF1 was spontaneously down-regulated after d 1-3 of neural induction. By double-knockdown experiments, up-regulation of SOX2 was critical for the increase of PAX6 and SOX1 expression in shRNA targeting Atf1 hESCs. Using the luciferase reporter assay, we identified ATF1 as a negative transcriptional regulator of Sox2 gene expression. A novel function of ATF1 was discovered, and these findings contribute to a broader understanding of the very first steps in regulating NE differentiation in hESCs.-Yang, S.-C., Liu, J.-J., Wang, C.-K., Lin, Y.-T., Tsai, S.-Y., Chen, W.-J., Huang, W.-K., Tu, P.-W. A., Lin, Y.-C., Chang, C.-F., Cheng, C.-L., Lin, H., Lai, C.-Y., Lin, C.-Y., Lee, Y.-H., Chiu, Y.-C., Hsu, C.-C., Hsu, S.-C., Hsiao, M., Schuyler, S. C., Lu, F. L., Lu, J. Down-regulation of ATF1 leads to early neuroectoderm differentiation of human embryonic stem cells by increasing the expression level of SOX2.

Keywords: early development; gatekeeper; high-throughput screen.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Activating Transcription Factor 1 / antagonists & inhibitors
  • Activating Transcription Factor 1 / genetics
  • Activating Transcription Factor 1 / metabolism*
  • Cell Differentiation*
  • Cells, Cultured
  • Down-Regulation
  • Endoderm / cytology
  • Endoderm / metabolism
  • Gene Expression Regulation, Developmental*
  • Human Embryonic Stem Cells / cytology*
  • Human Embryonic Stem Cells / metabolism
  • Humans
  • Mesoderm / cytology
  • Mesoderm / metabolism
  • Neurons / cytology*
  • Neurons / metabolism
  • RNA, Small Interfering / genetics*
  • SOXB1 Transcription Factors / genetics
  • SOXB1 Transcription Factors / metabolism*

Substances

  • ATF1 protein, human
  • Activating Transcription Factor 1
  • RNA, Small Interfering
  • SOX2 protein, human
  • SOXB1 Transcription Factors