Generation of INS-jGCaMP7f knock-in Ca(2+) reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based gene targeting
Summary
As a member of the single-fluorophore genetically encoded calcium indicators (GECIs), jGCaMP7f is widely applied to investigate intracellular Ca2+ concentrations. Here, we established an INS-jGCaMP7f knock-in H1 human embryonic stem cell (hESC) line by integrating jGCaMP7f gene into insulin locus via CRISPR/Cas9 system. The reporter cell line not only effectively labelled the insulin-producing cells induced from hESC, but also reflected the cytosolic change of Ca2+ level in response to different stimuli. This reporter cell line is a valuable research tool for studying functional maturation of hESC-derived insulin-producing cells, conducting drug screenings, and exploring the mechanisms of diabetes. Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.
| Authors | Liu X, Zhang F, Chen D, Yin J, Bi Z, Chen L, Yan J, Dong Q, Peng W, Xu T, Guo Y, Lin H, Liu H |
|---|---|
| Journal | Stem cell research |
| Publication Date | 2025 Feb;82:103633 |
| PubMed | 39708407 |
| DOI | 10.1016/j.scr.2024.103633 |