Zfp322a Regulates mouse ES cell pluripotency and enhances reprogramming efficiency

PLoS Genet. 2014 Feb 13;10(2):e1004038. doi: 10.1371/journal.pgen.1004038. eCollection 2014 Feb.

Abstract

Embryonic stem (ES) cells derived from the inner cell mass (ICM) of blastocysts are characterised by their ability to self-renew and their potential to differentiate into many different cell types. Recent studies have shown that zinc finger proteins are crucial for maintaining pluripotent ES cells. Mouse zinc finger protein 322a (Zfp322a) is expressed in the ICM of early mouse embryos. However, little is known regarding the role of Zfp322a in the pluripotency maintenance of mouse ES cells. Here, we report that Zfp322a is required for mES cell identity since depletion of Zfp322a directs mES cells towards differentiation. Chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assays revealed that Zfp322a binds to Pou5f1 and Nanog promoters and regulates their transcription. These data along with the results obtained from our ChIP-seq experiment showed that Zfp322a is an essential component of mES cell transcription regulatory network. Targets which are directly regulated by Zfp322a were identified by correlating the gene expression profile of Zfp322a RNAi-treated mES cells with the ChIP-seq results. These experiments revealed that Zfp322a inhibits mES cell differentiation by suppressing MAPK pathway. Additionally, Zfp322a is found to be a novel reprogramming factor that can replace Sox2 in the classical Yamanaka's factors (OSKM). It can be even used in combination with Yamanaka's factors and that addition leads to a higher reprogramming efficiency and to acceleration of the onset of the reprogramming process. Together, our results demonstrate that Zfp322a is a novel essential component of the transcription factor network which maintains the identity of mouse ES cells.

MeSH terms

  • Animals
  • Cell Differentiation / genetics*
  • Cellular Reprogramming / genetics*
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism*
  • Gene Expression Regulation, Developmental
  • Mice
  • Octamer Transcription Factor-3
  • Pluripotent Stem Cells / cytology
  • Pluripotent Stem Cells / metabolism*
  • Promoter Regions, Genetic
  • Transcription Factors / genetics*
  • Zinc Fingers / genetics

Substances

  • DNA-Binding Proteins
  • Octamer Transcription Factor-3
  • Transcription Factors
  • Zfp322a protein, mouse

Grants and funding

This work was supported by Singapore Ministry of Education and Singapore National Medical Research Council. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.