利用 CRISPR/Cas9 编辑技术建立 CPAMD8-GFP 报告人类胚胎干细胞系 IBBDe001-B

IF 0.8 4区医学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Stem cell research Pub Date : 2024-11-22 DOI:10.1016/j.scr.2024.103615

Shun Zhang , Jiahang Li , Wenmin Pan , Qing Ren , Yao Zhou , Ming Chen , Jia Qu , Shasha Li

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引用次数: 0

摘要

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，主要表达于形成虹膜和睫状体的视网膜神经上皮的远端。CPAMD8 基因突变与前节发育不良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 3*EAAAK-EGFP 荧光标签，从而能够实时观察其表达情况，并深入了解其生物学功能。由此产生的基因编辑细胞系保留了正常的干细胞形态和核型，表达了基本的多能性标记，并表现出分化潜能。

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Establishment of a CPAMD8-GFP reporter human embryonic stem cell line, IBBDe001-B, using CRISPR/Cas9 editing

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.

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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.