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

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

doi:10.1016/j.scr.2024.103615

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

Stem Cell Res. 2024 Nov 22:81:103615. doi: 10.1016/j.scr.2024.103615. Online ahead of print.

Authors

Shun Zhang¹, Jiahang Li², Wenmin Pan¹, Qing Ren², Yao Zhou², Ming Chen², Jia Qu³, Shasha Li⁴

Affiliations

¹ 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.
² National Engineering Research Center of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, China.
³ 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; Oujiang Laboratory, Zhejiang Lab for Regenerative Medicine, Vision and Brain Health, Wenzhou, China. Electronic address: qujia@eye.ac.cn.
⁴ 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. Electronic address: shashali2022@126.com.

PMID: 39603092
DOI: 10.1016/j.scr.2024.103615

Abstract

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.