CRISPR/Cas9-mediated targeted gene correction in amyotrophic lateral sclerosis patient iPSCs

Lixia Wang; Fei Yi; Lina Fu; Jiping Yang; Si Wang; Zhaoxia Wang; Keiichiro Suzuki; Liang Sun; Xiuling Xu; Yang Yu; Jie Qiao; Juan Carlos Izpisua Belmonte; Ze Yang; Yun Yuan; Jing Qu; Guang-Hui Liu

doi:10.1007/s13238-017-0397-3

CRISPR/Cas9-mediated targeted gene correction in amyotrophic lateral sclerosis patient iPSCs

Protein Cell. 2017 May;8(5):365-378. doi: 10.1007/s13238-017-0397-3. Epub 2017 Apr 11.

Authors

Lixia Wang^{1

2

3}, Fei Yi⁴, Lina Fu^{1

3}, Jiping Yang^{1

3}, Si Wang^{1

3}, Zhaoxia Wang⁵, Keiichiro Suzuki^{6

7}, Liang Sun⁸, Xiuling Xu¹, Yang Yu⁹, Jie Qiao⁹, Juan Carlos Izpisua Belmonte⁶, Ze Yang⁸, Yun Yuan¹⁰, Jing Qu^{11

12}, Guang-Hui Liu^{13

14

15

16}

Affiliations

¹ National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China.
² State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China.
³ University of Chinese Academy of Sciences, Beijing, 100049, China.
⁴ Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA, 94305, USA.
⁵ Department of Neurology, Peking University First Hospital, Beijing, 100034, China.
⁶ Gene Expression Laboratory, Salk Institute for Biological Studies, 10010 North Torrey Pines Road, La Jolla, CA, 92037, USA.
⁷ Universidad Católica San Antonio de Murcia (UCAM), Campus de los Jerónimos, N 135 Guadalupe, 30107, Murcia, Spain.
⁸ Beijing Hospital of the Ministry of Health, Beijing, 100730, China.
⁹ Department of Gynecology and Obstetrics, Peking University Third Hospital, Beijing, 100191, China.
¹⁰ Department of Neurology, Peking University First Hospital, Beijing, 100034, China. yuanyun2002@126.com.
¹¹ State Key Laboratory of Stem Cell and Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, 100101, China. qujing@ioz.ac.cn.
¹² University of Chinese Academy of Sciences, Beijing, 100049, China. qujing@ioz.ac.cn.
¹³ National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, China. ghliu@ibp.ac.cn.
¹⁴ University of Chinese Academy of Sciences, Beijing, 100049, China. ghliu@ibp.ac.cn.
¹⁵ Key Laboratory of Regenerative Medicine of Ministry of Education, Institute of Aging and Regenerative Medicine, Jinan University, Guangzhou, 510632, China. ghliu@ibp.ac.cn.
¹⁶ Beijing Institute for Brain Disorders, Capital Medical University, Beijing, 100069, China. ghliu@ibp.ac.cn.

Abstract

Amyotrophic lateral sclerosis (ALS) is a complex neurodegenerative disease with cellular and molecular mechanisms yet to be fully described. Mutations in a number of genes including SOD1 and FUS are associated with familial ALS. Here we report the generation of induced pluripotent stem cells (iPSCs) from fibroblasts of familial ALS patients bearing SOD1 ^+/A272C and FUS ^+/G1566A mutations, respectively. We further generated gene corrected ALS iPSCs using CRISPR/Cas9 system. Genome-wide RNA sequencing (RNA-seq) analysis of motor neurons derived from SOD1 ^+/A272C and corrected iPSCs revealed 899 aberrant transcripts. Our work may shed light on discovery of early biomarkers and pathways dysregulated in ALS, as well as provide a basis for novel therapeutic strategies to treat ALS.

Keywords: ALS; CRISPR/Cas9; gene correction; iPSC disease modeling.

Publication types

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

MeSH terms

Amyotrophic Lateral Sclerosis* / genetics
Amyotrophic Lateral Sclerosis* / metabolism
Amyotrophic Lateral Sclerosis* / therapy
Cell Line
Clustered Regularly Interspaced Short Palindromic Repeats*
Genetic Therapy*
Genome-Wide Association Study
Humans
Induced Pluripotent Stem Cells / metabolism*
Mutation, Missense
RNA-Binding Protein FUS* / genetics
RNA-Binding Protein FUS* / metabolism
Superoxide Dismutase-1* / genetics
Superoxide Dismutase-1* / metabolism

Substances

FUS protein, human
RNA-Binding Protein FUS
SOD1 protein, human
Superoxide Dismutase-1