Generation of INS-jGCaMP7f knock-in Ca2+ reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based gene targeting
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 vi...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-02-01
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| Series: | Stem Cell Research |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S1873506124003313 |
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| author | Xin Liu Feng Zhang Deqi Chen Jiaxiang Yin Zirong Bi Lihua Chen Jiawei Yan Qifei Dong Wei Peng Tao Xu Yanying Guo Haopeng Lin Huisheng Liu |
| author_facet | Xin Liu Feng Zhang Deqi Chen Jiaxiang Yin Zirong Bi Lihua Chen Jiawei Yan Qifei Dong Wei Peng Tao Xu Yanying Guo Haopeng Lin Huisheng Liu |
| author_sort | Xin Liu |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-c41ecd1aed2b478a957b0de41dfbe198 |
| institution | OA Journals |
| issn | 1873-5061 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Research |
| spelling | doaj-art-c41ecd1aed2b478a957b0de41dfbe1982025-08-20T02:35:47ZengElsevierStem Cell Research1873-50612025-02-018210363310.1016/j.scr.2024.103633Generation of INS-jGCaMP7f knock-in Ca2+ reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based gene targetingXin Liu0Feng Zhang1Deqi Chen2Jiaxiang Yin3Zirong Bi4Lihua Chen5Jiawei Yan6Qifei Dong7Wei Peng8Tao Xu9Yanying Guo10Haopeng Lin11Huisheng Liu12Department 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, ChinaGuangzhou National Laboratory, Guangzhou 510005, ChinaGuangzhou National Laboratory, Guangzhou 510005, ChinaGuangzhou National Laboratory, Guangzhou 510005, China; School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, ChinaGuangzhou National Laboratory, Guangzhou 510005, ChinaGuangzhou National Laboratory, Guangzhou 510005, China; School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, ChinaGuangzhou National Laboratory, Guangzhou 510005, China; School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, ChinaGuangzhou National Laboratory, Guangzhou 510005, China; Institute of Cytology and Genetics, School of Basic Medical Sciences, Hengyang Medical School, University of South China, Hengyang, Hunan 421001, ChinaGuangzhou National Laboratory, Guangzhou 510005, China; Institute of Cytology and Genetics, School of Basic Medical Sciences, Hengyang Medical School, University of South China, Hengyang, Hunan 421001, ChinaGuangzhou National Laboratory, Guangzhou 510005, China; School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, China; Corresponding authors.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; Corresponding authors.Guangzhou National Laboratory, Guangzhou 510005, China; School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, China; Corresponding authors.Guangzhou National Laboratory, Guangzhou 510005, China; School of Biomedical Engineering, Guangzhou Medical University, Guangzhou 510180, China; Bioland Laboratory, Guangzhou 510005, China; Corresponding authors.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.http://www.sciencedirect.com/science/article/pii/S1873506124003313jGCaMP7fCa2+InsulinhESCCRISPR/Cas9 |
| spellingShingle | Xin Liu Feng Zhang Deqi Chen Jiaxiang Yin Zirong Bi Lihua Chen Jiawei Yan Qifei Dong Wei Peng Tao Xu Yanying Guo Haopeng Lin Huisheng Liu Generation of INS-jGCaMP7f knock-in Ca2+ reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based gene targeting Stem Cell Research jGCaMP7f Ca2+ Insulin hESC CRISPR/Cas9 |
| title | Generation of INS-jGCaMP7f knock-in Ca2+ reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based gene targeting |
| title_full | Generation of INS-jGCaMP7f knock-in Ca2+ reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based gene targeting |
| title_fullStr | Generation of INS-jGCaMP7f knock-in Ca2+ reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based gene targeting |
| title_full_unstemmed | Generation of INS-jGCaMP7f knock-in Ca2+ reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based gene targeting |
| title_short | Generation of INS-jGCaMP7f knock-in Ca2+ reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based gene targeting |
| title_sort | generation of ins jgcamp7f knock in ca2 reporter human embryonic stem cell line gzle001 c using crispr cas9 based gene targeting |
| topic | jGCaMP7f Ca2+ Insulin hESC CRISPR/Cas9 |
| url | http://www.sciencedirect.com/science/article/pii/S1873506124003313 |
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