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Updated: May 3, 2026
Zinc-finger Nuclease Enhanced Gene Targeting in Human Embryonic Stem Cells
Published on: August 23, 2014
Generation of INS-jGCaMP7f knock-in Ca2+ reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based
Xin Liu1, Feng Zhang2, Deqi Chen2
1Department 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.
Abstract:
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.
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