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

Authors :
Daniele Merico
Karun K. Singh
James Ellis
Peter Pasceri
Jennifer L. Howe
Deivid C. Rodrigues
Gaganjot Kaur
Ryan K. C. Yuen
Bhooma Thiruvahindrapuram
Susan Walker
P. Joel Ross
Alina Piekna
Sean H. White
Roumiana Alexandrova
Giovanna Pellecchia
Zhuozhi Wang
Eric Deneault
Stephen W. Scherer
Kirill Zaslavsky
Vickie Kwan
Wei Wei
Muhammad Faheem
Publication Year :
2018
Publisher :
Cold Spring Harbor Laboratory, 2018.

Abstract

SummaryAutism Spectrum Disorder is phenotypically and genetically heterogeneous, but genomic analyses have identified candidate susceptibility genes. 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 10 ASD-relevant genes (AFF2/FMR2, ANOS1, ASTN2, ATRX, CACNA1C, CHD8, DLGAP2, KCNQ2, SCN2A, TENM1). Neurogenin 2 (NEUROG2)-directed differentiation of iPSCs allowed production of cortical excitatory neurons, and mutant proteins were not detectable. RNAseq 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 post-synaptic 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.

Details

Language :
English
Database :
OpenAIRE
Accession number :
edsair.doi.dedup.....9ae79822ea856a786554f34ca2e06b0e
Full Text :
https://doi.org/10.1101/344234