Introduction of a Geminin mScarlet Reporter into H2B-mTurq2 hiPSCs for Live-cell Imaging of Proliferation and Cell Cycling


We previously generated a doxycycline-inducible H2B-mTurq2 reporter in hiPSCs to track cells and study cell division and apoptosis. To improve visualization of cycling cells, we introduced a ubiquitously transcribed mScarletI-Geminin (GMMN) (1-110) into the previously untargeted second AAVS1 allele. Fusion to the N-terminal part of GMNN provided tightly controlled mScarletI expression during the cell cycle. mScarletI fluorescence increased gradually from the S-phase through the M-phase of the cell cycle and was lost at the metaphase-anaphase transition. The resulting hiPSC reporter line generated, which we named ProLiving, is a valuable tool to study cell division and cell cycle characteristics in living hiPSC-derived cells. Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.

Authors Arendzen CH, Cramer SJ, Freund CMAH, Mummery CL, Ranga A, Mikkers HMM
Journal Stem cell research
Publication Date 2023 Mar;67:103031
PubMed 36702081
DOI 10.1016/j.scr.2023.103031

Research Projects

Cell Lines