通过CRISPR/Cas9编辑生成BEST1 Pr-EGFP报告基因人胚胎干细胞系。
Generation of a BEST1 Pr-EGFP reporter human embryonic stem cell line via CRISPR/Cas9 editing.
作者信息
Lu Shuaiyan, Chen Ming, Liu Xiaoyu, Li Jiahang, Liu Hui, Li Shasha
机构信息
National Engineering Research Center of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, China; Oujiang Laboratory (Zhejiang Lab for Regenerative Medicine, Vision and Brain Health), Wenzhou, China; State Key Laboratory of Ophthalmology, Optometry and Visual Science, Eye Hospital, Wenzhou Medical University, Wenzhou, China.
National Engineering Research Center of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, China; State Key Laboratory of Ophthalmology, Optometry and Visual Science, Eye Hospital, Wenzhou Medical University, Wenzhou, China.
出版信息
Stem Cell Res. 2025 Feb;82:103625. doi: 10.1016/j.scr.2024.103625. Epub 2024 Dec 9.
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.
视网膜色素上皮（RPE）细胞位于神经视网膜和脉络膜毛细血管之间，对视网膜维持和光感受器功能至关重要。人类胚胎干细胞（hESCs）为移植提供了无限的RPE细胞来源。我们使用CRISPR/Cas9将BEST1启动子（一种RPE特异性标志物）与EGFP基因的融合体插入AAVS1位点，以追踪hESC诱导的RPE（hESC-iRPE）中的分化情况。产生的基因编辑系WAe009-A-2 M保持正常核型，表达多能性标志物，并表现出分化潜能，这使其在RPE发育和治疗研究中具有极高价值。
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