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Arendzen, C.H.; Chaudhari, U.; Cramer, S.J.; Freund, C.M.A.H.; Mummery, C.L.; Ranga, A.; Pourquie, O.; Mikkers, H.M.M.
Stem cell research, December 2021, 2021-Dec, 2021-12-00, 2021-12-01, Volume: 57Journal Article
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.
