Stem Cell Research (Dec 2021)

Generation of LUMCi041-A-2: Equipping a PAX3 reporter iPSC line with doxycycline inducible H2B-mTurquoise2 for live cell imaging

  • C.H. Arendzen,
  • U. Chaudhari,
  • S.J. Cramer,
  • C.M.A.H. Freund,
  • C.L. Mummery,
  • A. Ranga,
  • O. Pourquie,
  • H.M.M. Mikkers

Journal volume & issue
Vol. 57
p. 102592

Abstract

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An induced pluripotent stem cell (iPSC) line, in which a H2B-fluorescent protein fusion is temporally expressed, is a valuable tool to track cells and study cell divisions and apoptosis. To this end we introduced a 3rd generation “all-in-one” doxycycline-inducible H2B-mTurquoise2 vector into the AAVS1 locus of PAX3-Venus iPSCs via CRISPR/Cas9. H2B-mTurquoise2 expression is absent but readily induced by doxycycline allowing quantification of cell divisions and imaging of living cells. Besides being a universal reporter in iPSC-based differentiation and toxicity assays, the generated pluripotent and genomically normal LUMCi041-A-2 line is particularly suited to study PAX3-positive stages of development.

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