利用CRISPR/Cas9编辑技术建立CPAMD8-GFP报告基因人胚胎干细胞系IBBDe001-B。
Establishment of a CPAMD8-GFP reporter human embryonic stem cell line, IBBDe001-B, using CRISPR/Cas9 editing.
作者信息
Zhang Shun, Li Jiahang, Pan Wenmin, Ren Qing, Zhou Yao, Chen Ming, Qu Jia, Li Shasha
机构信息
School of Biomedical Engineering, Hainan University, Haikou, China; National Engineering Research Center of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, China; Hainan Institute of Real World Data, Qionghai, China.
National Engineering Research Center of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, China.
出版信息
Stem Cell Res. 2024 Dec;81:103615. doi: 10.1016/j.scr.2024.103615. Epub 2024 Nov 22.
The human CPAMD8 gene encodes proteins in the A2M/C3 (alpha-2-macroglobulin/complement 3) family, predominantly expressed in the distal tips of the retinal neuroepithelium that form the iris and ciliary body. Mutations in CPAMD8 have been linked to anterior segment dysplasia and congenital glaucoma. Using CRISPR/Cas9 editing, we inserted a 3*EAAAK-EGFP fluorescent tag into the CPAMD8 gene, enabling real-time observation of its expression and providing insights into its biological functions. The resulting gene-edited cell line retained normal stem cell morphology and karyotype, expressed essential pluripotency markers, and exhibited differentiation potential.
人类CPAMD8基因编码A2M/C3（α-2-巨球蛋白/补体3）家族中的蛋白质，主要在形成虹膜和睫状体的视网膜神经上皮的远端尖端表达。CPAMD8基因的突变与前段发育异常和先天性青光眼有关。利用CRISPR/Cas9编辑技术，我们在CPAMD8基因中插入了一个3*EAAAK-EGFP荧光标签，能够实时观察其表达情况，并深入了解其生物学功能。所得的基因编辑细胞系保留了正常的干细胞形态和核型，表达重要的多能性标志物，并具有分化潜能。
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