Complete Disruption of Autism-Susceptibility Genes by Gene Editing Predominantly Reduces Functional Connectivity of Isogenic Human Neurons

被引:89
作者
Deneault, Eric [1 ,2 ]
White, Sean H. [3 ]
Rodrigues, Deivid C. [4 ]
Ross, Joel [4 ,7 ]
Faheem, Muhammad [1 ,2 ]
Zaslavsky, Kirill [4 ,5 ]
Wang, Zhuozhi [2 ]
Alexandrova, Roumiana [2 ]
Pellecchia, Giovanna [2 ]
Wei, Wei [4 ]
Piekna, Alina [4 ]
Kaur, Gaganjot [2 ]
Howe, Jennifer L. [2 ]
Kwan, Vickie [3 ]
Thiruvahindrapuram, Bhooma [2 ]
Walker, Susan [1 ,2 ]
Pasceri, Peter [4 ]
Merico, Daniele [2 ,8 ]
Yuen, Ryan Kc [1 ,2 ]
Singh, Karun K. [3 ]
Ellis, James [4 ,5 ]
Scherer, Stephen W. [1 ,2 ,5 ,6 ]
机构
[1] Hosp Sick Children, Genet & Genome Biol Program, Toronto, ON M5G 1X8, Canada
[2] Hosp Sick Children, Ctr Appl Genom, Toronto, ON M5G 1X8, Canada
[3] McMaster Univ, Dept Biochem & Biomed Sci, Stem Cell & Canc Res Inst, Hamilton, ON L8S 4L8, Canada
[4] Hosp Sick Children, Dev & Stem Cell Biol Program, Toronto, ON M5G 1X8, Canada
[5] Univ Toronto, Dept Mol Genet, Toronto, ON M5S 3H7, Canada
[6] Univ Toronto, McLaughlin Ctr, Toronto, ON M5S 3H7, Canada
[7] Univ Prince Edward Isl, Dept Biol, Charlottetown, PE C1A 4P3, Canada
[8] Deep Genom Inc, Toronto, ON M5G 1M1, Canada
来源
STEM CELL REPORTS | 2018年 / 11卷 / 05期
基金
加拿大创新基金会; 加拿大自然科学与工程研究理事会; 加拿大健康研究院;
关键词
SPECTRUM DISORDER; BINDING PROTEIN; RISK; SCHIZOPHRENIA; ASSOCIATION; RECURRENCE; INDUCTION; VARIANTS; SIBLINGS; MUTATION;
D O I
10.1016/j.stemcr.2018.10.003
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Autism spectrum disorder (ASD) is phenotypically and genetically heterogeneous. We present a CRISPR gene editing strategy to insert a protein tag and premature termination sites creating an induced pluripotent stem cell (iPSC) knockout resource for functional studies of ten ASD-relevant genes (AFF2/FMR2, ANOS1, ASTN2, ATRX, CACNA1C, CHD8, DLGAP2, KCNQ2, SCN2A, TENM1). Neurogenin 2 (NGN2)-directed induction of iPSCs allowed production of excitatory neurons, and mutant proteins were not detectable. RNA sequencing revealed convergence of several neuronal networks. Using both patch-clamp and multi-electrode array approaches, the electrophysiological deficits measured were distinct for different mutations. However, they culminated in a consistent reduction in synaptic activity, including reduced spontaneous excitatory postsynaptic current frequencies in AFF2/FMR2-, ASTN2-, ATRX-, KCNQ2-, and SCN2A-null neurons. Despite ASD susceptibility genes belonging to different gene ontologies, isogenic stem cell resources can reveal common functional phenotypes, such as reduced functional connectivity.
引用
收藏
页码:1211 / 1225
页数:15
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