Generation of a DKK2 homozygous knockout human embryonic stem cell line using the CRISPR/Cas9 system

Wang Zhang; Yuming Li; Li Wang; Tao Xin; Canwei Zhang

Stem Cell Research (Dec 2021)

Generation of a DKK2 homozygous knockout human embryonic stem cell line using the CRISPR/Cas9 system

Wang Zhang,
Yuming Li,
Li Wang,
Tao Xin,
Canwei Zhang

Affiliations

Wang Zhang: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China
Yuming Li: Department of Neurosurgery, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Shandong Medicine and Health Key Laboratory of Neurosurgery, Jinan 250014, China
Li Wang: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China
Tao Xin: Department of Neurosurgery, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Shandong Medicine and Health Key Laboratory of Neurosurgery, Jinan 250014, China; Corresponding authors at: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China (C. Zhang).
Canwei Zhang: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China; Department of Ophthalmology, The First Affiliated Hospital of Shandong First Medical University & Shandong Provincial Qianfoshan Hospital, Jinan 250014, China; Corresponding authors at: State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou 510060, China (C. Zhang).

Journal volume & issue: Vol. 57
p. 102611

Abstract

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Dickkopf-2 (DKK2) is an antagonist of canonical Wnt signaling, which is involved in various biological processes of development, such as epidermal appendage formation and eye development. To identify underlying effects of DKK2 during embryonic development, we generated a DKK2 homozygous knockout human embryonic stem cell (hESC) line through the CRISPR/Cas9 genome-editing technology. This cell line, which maintains normal stem cell morphology and stably expresses pluripotent markers, could provide an ideal platform for exploring the role of DKK2 in embryonic development. In addition, Zeocin selection combined with tiny clone picking might be a highly efficient way to generate gene-knockout hESC lines.

Published in Stem Cell Research

ISSN: 1873-5061 (Print); 1876-7753 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Science: Biology (General)
Website: https://www.journals.elsevier.com/stem-cell-research

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