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

Summary

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. Copyright © 2021 The Authors. Published by Elsevier B.V. All rights reserved.

Authors Arendzen CH, Chaudhari U, Cramer SJ, Freund CMAH, Mummery CL, Ranga A, Pourquie O, Mikkers HMM
Journal Stem cell research
Publication Date 2021 Nov 3;57:102592
PubMed 34775202
DOI 10.1016/j.scr.2021.102592

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