利用基于CRISPR/Cas9的基因靶向技术生成INS-jGCaMP7f敲入型钙报告基因人胚胎干细胞系GZLe001-C。
Generation of INS-jGCaMP7f knock-in Ca reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based gene targeting.
作者信息
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
机构信息
Department of Endocrinology and Metabolism, People's Hospital of Xinjiang Uygur Autonomous Region, Xinjiang Clinical Research Center for Diabetes, Xinjiang Key Laboratory of Cardiovascular Homeostasis and Regeneration Research, Urumqi 830001, China; Guangzhou National Laboratory, Guangzhou 510005, China.
Guangzhou National Laboratory, Guangzhou 510005, China.
出版信息
Stem Cell Res. 2025 Feb;82:103633. doi: 10.1016/j.scr.2024.103633. Epub 2024 Dec 18.
As a member of the single-fluorophore genetically encoded calcium indicators (GECIs), jGCaMP7f is widely applied to investigate intracellular Ca 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 Ca 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.
作为单荧光团基因编码钙指示剂（GECIs）的一员，jGCaMP7f被广泛应用于研究细胞内钙浓度。在此，我们通过CRISPR/Cas9系统将jGCaMP7f基因整合到胰岛素基因座，建立了INS-jGCaMP7f敲入H1人胚胎干细胞（hESC）系。该报告细胞系不仅能有效标记由hESC诱导产生的胰岛素分泌细胞，还能反映不同刺激下细胞溶质钙水平的变化。该报告细胞系是研究hESC来源的胰岛素分泌细胞功能成熟、进行药物筛选以及探索糖尿病发病机制的宝贵研究工具。
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