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利用 CRISPR/Cas9 生成并鉴定 GATA6 特异性 EGFP 表达的人诱导多能干细胞系,KSCBi017-A-1。

Generation and characterization of GATA6-specific EGFP expressing human induced pluripotent stem cell line, KSCBi017-A-1, using CRISPR/Cas9.

机构信息

Center for Genomics and Proteomics & Stem Cell Core Facility, Lee Gil Ya Cancer and Diabetes Institute, Gachon University, Incheon, Republic of Korea.

Division of Intractable Disease Research, Korea National Institute of Health, Osong, Cheongju, Republic of Korea.

出版信息

Stem Cell Res. 2024 Jun;77:103426. doi: 10.1016/j.scr.2024.103426. Epub 2024 Apr 24.

Abstract

GATA6 is expressed during early embryogenesis and localizes to endoderm- and mesoderm-derived tissues during later embryogenesis. Here, we established a human induced pluripotent stem cell (hiPSC) line expressing EGFP under GATA6 gene. EGFP coding sequence was introduced into the C-terminus of GATA6 in KSCBi017-A hiPSCs through homologous recombination using CRISPR/Cas9 system. The successfully edited line, KSCBi017-A-1, was selected and confirmed by sequencing. The line had a normal karyotype and exhibited potential to differentiate into three germ layers while it expressed EGFP upon endoderm induction. KSCBi017-A-1 cells can be used to monitor the expression of GATA6 during differentiation. This cell line is available from Korea National Stem Cell Bank.

摘要

GATA6 在胚胎早期表达,并在胚胎后期定位于内胚层和中胚层来源的组织。在这里,我们建立了一条表达 EGFP 的人诱导多能干细胞(hiPSC)系,该基因受 GATA6 调控。通过 CRISPR/Cas9 系统,将 EGFP 编码序列插入 KSCBi017-A hiPSCs 的 GATA6 基因 C 末端。通过同源重组成功编辑的细胞系 KSCBi017-A-1 通过测序进行了筛选和验证。该细胞系具有正常的核型,并表现出向三个胚层分化的潜能,同时在内胚层诱导时表达 EGFP。KSCBi017-A-1 细胞可用于监测分化过程中 GATA6 的表达。该细胞系可从韩国国家干细胞库获得。

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