Generation of INS-jGCaMP7f knock-in Ca2+ reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based gene targeting

• Published in: Stem cell research 2024-12, Vol.82, p.103633
• Main Authors: Liu, Xin, Zhang, Feng, Chen, Deqi, Yin, Jiaxiang, Bi, Zirong, Chen, Lihua, Yan, Jiawei, Dong, Qifei, Peng, Wei, Xu, Tao, Guo, Yanying, Lin, Haopeng, Liu, Huisheng
• Format: Article
• Language: English
• ISSN: 1876-7753; 1876-7753
• DOI: 10.1016/j.scr.2024.103633

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.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.