利用CRISPR/ cas9基因靶向技术生成INS-jGCaMP7f敲入Ca2+报告基因的人胚胎干细胞系GZLe001-C

IF 0.8 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Stem cell research Pub Date : 2025-02-01 DOI:10.1016/j.scr.2024.103633

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

{"title":"利用CRISPR/ cas9基因靶向技术生成INS-jGCaMP7f敲入Ca2+报告基因的人胚胎干细胞系GZLe001-C","authors":"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","doi":"10.1016/j.scr.2024.103633","DOIUrl":null,"url":null,"abstract":"<div><div>As a member of the single-fluorophore genetically encoded calcium indicators (GECIs), jGCaMP7f is widely applied to investigate intracellular Ca<sup>2+</sup> 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<sup>2+</sup> 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.</div></div>","PeriodicalId":21843,"journal":{"name":"Stem cell research","volume":"82 ","pages":"Article 103633"},"PeriodicalIF":0.8000,"publicationDate":"2025-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Generation of INS-jGCaMP7f knock-in Ca2+ reporter human embryonic stem cell line, GZLe001-C, using CRISPR/Cas9-based gene targeting\",\"authors\":\"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\",\"doi\":\"10.1016/j.scr.2024.103633\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>As a member of the single-fluorophore genetically encoded calcium indicators (GECIs), jGCaMP7f is widely applied to investigate intracellular Ca<sup>2+</sup> 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<sup>2+</sup> 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.</div></div>\",\"PeriodicalId\":21843,\"journal\":{\"name\":\"Stem cell research\",\"volume\":\"82 \",\"pages\":\"Article 103633\"},\"PeriodicalIF\":0.8000,\"publicationDate\":\"2025-02-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Stem cell research\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1873506124003313\",\"RegionNum\":4,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"BIOTECHNOLOGY & APPLIED MICROBIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Stem cell research","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1873506124003313","RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOTECHNOLOGY & APPLIED MICROBIOLOGY","Score":null,"Total":0}

引用次数: 0

摘要

jGCaMP7f作为单荧光团基因编码钙指标（GECIs）的一员，被广泛应用于细胞内Ca2+浓度的研究。本研究通过CRISPR/Cas9系统将jGCaMP7f基因整合到胰岛素位点，建立了胰岛素-jGCaMP7f敲入H1人胚胎干细胞（hESC）细胞系。报告细胞系不仅能有效标记hESC诱导的胰岛素生成细胞，还能反映细胞内Ca2+水平在不同刺激下的变化。该报告细胞系是研究hesc来源的胰岛素生成细胞功能成熟、进行药物筛选和探索糖尿病机制的重要研究工具。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

查看原文本刊更多论文

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 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.

求助全文

通过发布文献求助，成功后即可免费获取论文全文。去求助

来源期刊

Stem cell research 生物-生物工程与应用微生物

CiteScore

2.20

自引率

8.30%

发文量

338

审稿时长

55 days

期刊介绍： Stem Cell Research is dedicated to publishing high-quality manuscripts focusing on the biology and applications of stem cell research. Submissions to Stem Cell Research, may cover all aspects of stem cells, including embryonic stem cells, tissue-specific stem cells, cancer stem cells, developmental studies, stem cell genomes, and translational research. Stem Cell Research publishes 6 issues a year.