通过CRISPR/Cas9编辑生成BEST1 Pr-EGFP报告基因的人胚胎干细胞系

IF 0.8 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Stem cell research Pub Date : 2025-02-01 DOI:10.1016/j.scr.2024.103625

Shuaiyan Lu , Ming Chen , Xiaoyu Liu , Jiahang Li , Hui Liu , Shasha Li

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引用次数: 0

摘要

视网膜色素上皮细胞（RPE）位于神经视网膜和绒毛膜毛细血管之间，对视网膜的维持和光感受器功能至关重要。人胚胎干细胞（hESCs）为移植提供了无限的RPE细胞来源。利用CRISPR/Cas9，我们将BEST1启动子（一种RPE特异性标记）和EGFP基因的融合插入AAVS1位点，以跟踪hesc诱导的RPE （hESC-iRPE）的分化。由此产生的基因编辑系wae009 - a - 2m保持了正常的核型，表达了多能性标记，并显示出分化潜力，使其对RPE的发展和治疗研究具有宝贵的价值。

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Generation of a BEST1 Pr-EGFP reporter human embryonic stem cell line via CRISPR/Cas9 editing

The retinal pigment epithelium (RPE) cell, located between the neural retina and choriocapillaris, is vital for retinal maintenance and photoreceptor function. Human embryonic stem cells (hESCs) provide a limitless source of RPE cells for transplantation. Using CRISPR/Cas9, we inserted a fusion of the BEST1 promoter (an RPE-specific marker) and the EGFP gene into the AAVS1 locus to track differentiation in hESC-induced RPE (hESC-iRPE). The resulting gene-edited line, WAe009-A-2 M, maintained a normal karyotype, expressed pluripotency markers, and demonstrated differentiation potential, making it invaluable for RPE development and therapeutic research.

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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.