Generation of Nanog reporter mice that distinguish pluripotent stem cells from unipotent primordial germ cells

Genesis. 2019 Nov;57(11-12):e23334. doi: 10.1002/dvg.23334. Epub 2019 Sep 12.

Abstract

Nanog is a core transcription factor specifically expressed not only in the pluripotent stem cells (PSCs), such as embryonic stem cells (ESCs), embryonic germ cells (EGCs), and induced PSCs (iPSCs), but also in the unipotent primordial germ cells (PGCs). Although Nanog promoter/enhancer regions are well characterized by in vitro analyses, direct correlations between the regulatory elements for Nanog expression and in vivo expression patterns of Nanog have not been fully clarified. In this study, we generated Nanog-RFP transgenic (Tg) mice in which expression of red fluorescent protein (RFP) is driven by a 5.2 kb Nanog promoter/enhancer region. As expected, RFP was expressed in the inner cell mass of blastocysts, ESCs, and iPSCs. However, RFP fluorescence was not observed in PGCs, although Nanog was expressed in PGCs. Because RFP fluorescence was visible in the PGC-derived pluripotent EGCs in culture, it was suggested that the reporter gene expression was specifically activated in PSCs. In conclusion, we have generated a novel Nanog-RFP Tg mouse line that can selectively tag PSCs over unipotent PGCs.

Keywords: gene expression; pluripotency; transcription factor; transgenic mouse; unipotency.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cells, Cultured
  • Embryonic Stem Cells / metabolism
  • Germ Cells / metabolism
  • Homeodomain Proteins / genetics
  • Induced Pluripotent Stem Cells / metabolism
  • Mice
  • Mice, Transgenic
  • Nanog Homeobox Protein / metabolism*
  • Nanog Homeobox Protein / physiology
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / pathology*
  • Transcription Factors / genetics

Substances

  • Homeodomain Proteins
  • Nanog Homeobox Protein
  • Nanog protein, mouse
  • Transcription Factors