A stem cell reporter for investigating pluripotency and self-renewal in the rat.

Stephen Meek, Jun Wei, Taeho Oh, Jaime Olavarrieta, Linda Sutherland, Daniel F Carlson, Angela Salzano, Tamir Chandra, Anagha Joshi, Tom Burdon

Research output: Contribution to journalArticlepeer-review

Abstract

Rat embryonic stem cells (rESCs) are capable of contributing to all differentiated tissues including the germ line in chimaeric animals, and represent a unique, authentic, alternative to mouse ESCs for studying stem cell pluripotency and self-renewal. Here we describe an EGFP reporter transgene that tracks expression of the benchmark naïve pluripotency marker gene Rex1 (Zfp42) in the rat. Insertion of the EGFP reporter gene downstream of the Rex1 promoter
disrupted Rex1 expression, but REX1-deficient rESCs and rats were viable and apparently normal, validating this targeted knock-in transgene as a neutral reporter. The Rex1-EGFP gene responded to self-renewal/differentiation factors and validated the critical role of β- catenin/LEF1 signalling. The stem cell reporter also allowed the identification of functionally
distinct sub-populations of cells within rESC cultures, thus demonstrating its utility in discriminating between cell states in rat stem cell cultures, as well as providing a tool for tracking Rex1 expression in the rat.
Original languageEnglish
JournalStem Cell Reports
Early online date2 Jan 2020
DOIs
Publication statusE-pub ahead of print - 2 Jan 2020

Keywords

  • rat
  • embryonic stem cell
  • Rex1
  • Zfp42
  • fluorescent reporter
  • pluripotency
  • self-renewal
  • genetic engineering
  • transgenic

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