Generation of a tdTomato-GAD67 reporter human epilepsia mutation induced pluripotent stem cell line, USTCi001-A-2, using CRISPR/Cas9 editing

Huifang Zhao; Lang He; Hualin Huang; Shuai Li; Na Cheng; Feng Tang; Xiaobo Han; Zuoxian Lin; Rongqi Huang; Peng Zhou; Sihao Deng; Jufang Huang; Zhiyuan Li

doi:10.1016/j.scr.2020.102003

Generation of a tdTomato-GAD67 reporter human epilepsia mutation induced pluripotent stem cell line, USTCi001-A-2, using CRISPR/Cas9 editing

Stem Cell Res. 2020 Oct:48:102003. doi: 10.1016/j.scr.2020.102003. Epub 2020 Sep 17.

Authors

Huifang Zhao¹, Lang He², Hualin Huang³, Shuai Li³, Na Cheng⁴, Feng Tang³, Xiaobo Han³, Zuoxian Lin⁵, Rongqi Huang³, Peng Zhou⁴, Sihao Deng⁴, Jufang Huang⁴, Zhiyuan Li⁶

Affiliations

¹ School of Life Sciences, University of Science and Technology of China, Hefei, China; CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; Guangzhou Regenerative Medicine and Health Guangdong Laboratory, 510005 Guangzhou, China.
² Department of Neurology, Xiangya Hospital, Central South University, Changsha, Hunan, China.
³ CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; University of Chinese Academy of Sciences, Beijing 100049, China; Guangzhou Regenerative Medicine and Health Guangdong Laboratory, 510005 Guangzhou, China.
⁴ Department of Anatomy and Neurobiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China.
⁵ CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; Guangzhou Regenerative Medicine and Health Guangdong Laboratory, 510005 Guangzhou, China.
⁶ School of Life Sciences, University of Science and Technology of China, Hefei, China; CAS Key Laboratory of Regenerative Biology, Guangdong Provincial Key Laboratory of Stem Cell and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou 510530, China; CAS Key Laboratory of Regenerative Biology, Joint School of Life Sciences, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, 510530, China; Guangzhou Medical University, Guangzhou 511436, China; Department of Anatomy and Neurobiology, Xiangya School of Medicine, Central South University, Changsha, Hunan, China; University of Chinese Academy of Sciences, Beijing 100049, China; Guangzhou Regenerative Medicine and Health Guangdong Laboratory, 510005 Guangzhou, China. Electronic address: [email protected].

PMID: 32977294
DOI: 10.1016/j.scr.2020.102003

Abstract

Dravet syndrome is an epileptic encephalopathy largely due to haploinsufficiency of the voltage-gated sodium channel Nav1.1 that is expressed primarily in GABAergic neurons. In order to distinguish the different subtypes, we used gene editing to introduce tdTomato gene into the genome of iPSCs to label the GABAergic neurons in the differentiated neuronal networks. The gene-edited cell line demonstrates normal karyotype, expresses the main pluripotency markers, and shows the presence of differentiation into the three embryonic germ layers in teratomas.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

CRISPR-Cas Systems / genetics
Epilepsies, Myoclonic* / genetics
Humans
Induced Pluripotent Stem Cells*
Luminescent Proteins
Mutation / genetics
Red Fluorescent Protein

Substances

Luminescent Proteins