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....

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Main Authors: Shun Zhang, Jiahang Li, Wenmin Pan, Qing Ren, Yao Zhou, Ming Chen, Jia Qu, Shasha Li
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:Stem Cell Research
Online Access:http://www.sciencedirect.com/science/article/pii/S1873506124003131
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author Shun Zhang
Jiahang Li
Wenmin Pan
Qing Ren
Yao Zhou
Ming Chen
Jia Qu
Shasha Li
author_facet Shun Zhang
Jiahang Li
Wenmin Pan
Qing Ren
Yao Zhou
Ming Chen
Jia Qu
Shasha Li
author_sort Shun Zhang
collection DOAJ
description 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.
format Article
id doaj-art-31b3e7e4d1d842038b0cc304ea174374
institution OA Journals
issn 1873-5061
language English
publishDate 2024-12-01
publisher Elsevier
record_format Article
series Stem Cell Research
spelling doaj-art-31b3e7e4d1d842038b0cc304ea1743742025-08-20T01:59:30ZengElsevierStem Cell Research1873-50612024-12-018110361510.1016/j.scr.2024.103615Establishment of a CPAMD8-GFP reporter human embryonic stem cell line, IBBDe001-B, using CRISPR/Cas9 editingShun Zhang0Jiahang Li1Wenmin Pan2Qing Ren3Yao Zhou4Ming Chen5Jia Qu6Shasha Li7School 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, ChinaNational Engineering Research Center of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, ChinaSchool 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, ChinaNational Engineering Research Center of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, ChinaNational Engineering Research Center of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, ChinaNational Engineering Research Center of Ophthalmology and Optometry, Eye Hospital, Wenzhou Medical University, Wenzhou, ChinaSchool 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; Corresponding authors.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; Corresponding authors.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.http://www.sciencedirect.com/science/article/pii/S1873506124003131
spellingShingle Shun Zhang
Jiahang Li
Wenmin Pan
Qing Ren
Yao Zhou
Ming Chen
Jia Qu
Shasha Li
Establishment of a CPAMD8-GFP reporter human embryonic stem cell line, IBBDe001-B, using CRISPR/Cas9 editing
Stem Cell Research
title Establishment of a CPAMD8-GFP reporter human embryonic stem cell line, IBBDe001-B, using CRISPR/Cas9 editing
title_full Establishment of a CPAMD8-GFP reporter human embryonic stem cell line, IBBDe001-B, using CRISPR/Cas9 editing
title_fullStr Establishment of a CPAMD8-GFP reporter human embryonic stem cell line, IBBDe001-B, using CRISPR/Cas9 editing
title_full_unstemmed Establishment of a CPAMD8-GFP reporter human embryonic stem cell line, IBBDe001-B, using CRISPR/Cas9 editing
title_short Establishment of a CPAMD8-GFP reporter human embryonic stem cell line, IBBDe001-B, using CRISPR/Cas9 editing
title_sort establishment of a cpamd8 gfp reporter human embryonic stem cell line ibbde001 b using crispr cas9 editing
url http://www.sciencedirect.com/science/article/pii/S1873506124003131
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