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 Dec;57:102592 |
| PubMed | 34775202 |
| DOI | 10.1016/j.scr.2021.102592 |