Generation of a SOX2 reporter human induced pluripotent stem cell line using CRISPR/SaCas9

Stem Cell Res. 2017 Jul:22:16-19. doi: 10.1016/j.scr.2017.05.005. Epub 2017 May 17.

Abstract

SOX2 is an important transcription factor involved in pluripotency maintenance, pluripotent reprogramming and differentiation towards neural lineages. Here we engineered the previously described HEL24.3 hiPSC to generate a SOX2 reporter by knocking-in a T2A fused nuclear tdTomato reporter cassette before the STOP codon of the SOX2 gene coding sequence. CRISPR/SaCas9-mediated stimulation of homologous recombination was utilized to facilitate faithful targeted insertion. This line accurately reports the expression of endogenous SOX2 and therefore constitutes a useful tool to study the SOX2 expression dynamics upon hiPSC culture, differentiation and somatic cell reprogramming.

MeSH terms

  • CRISPR-Cas Systems*
  • Cell Differentiation / genetics
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism
  • Induced Pluripotent Stem Cells / physiology*
  • SOXB1 Transcription Factors / biosynthesis
  • SOXB1 Transcription Factors / genetics*

Substances

  • SOX2 protein, human
  • SOXB1 Transcription Factors