The generation and validation of two NKX2-5 fluorescent reporter human embryonic stem cell lines: UMANe002-A-1 and UMANe002-A-2

Stem Cell Res. 2024 Feb:74:103262. doi: 10.1016/j.scr.2023.103262. Epub 2023 Dec 3.

Abstract

The transcription factor NKX2-5 is a highly conserved master regulator of heart development which is widely expressed in cardiac progenitors and cardiomyocytes. Fluorescent reporters of NKX2-5 that minimally perturb normal protein expression can enable the identification, quantification and isolation of NKX2-5-expressing cells in a normal physiological state. Here we report the generation of two new hESC lines with eGFP inserted upstream (5') or downstream (3') of NKX2-5, linked by a cleavable T2A peptide. These complementary reporters produce a robust fluorescent signal in cardiac cells and have wide utility particularly for research on developmental biology and disease modelling.

Keywords: NKX2-5; cardiac; cardiomyocyte; pluripotent stem cell; progenitor; reporter.

MeSH terms

  • Cell Differentiation
  • Cell Line
  • Embryonic Stem Cells / metabolism
  • Homeobox Protein Nkx-2.5 / genetics
  • Homeobox Protein Nkx-2.5 / metabolism
  • Human Embryonic Stem Cells* / metabolism
  • Humans
  • Myocytes, Cardiac / metabolism

Substances

  • NKX2-5 protein, human
  • Homeobox Protein Nkx-2.5