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

• DOI: 10.1016/j.stemcr.2018.10.003
• Publication Date: 2018-11-01T14:43:49Z
• Authors: Eric Deneault; Sean H. White; Deivid C. Rodrigues; P. Joel Ross; Muhammad Faheem; Kirill Zaslavsky; Zhuozhi Wang; Roumiana Alexandrova; Giovanna Pellecchia; Wei Wei; Alina Piekna; Gaganjot Kaur; Jennifer L. Howe; Vickie Kwan; Bhooma Thiruvahin...; Susan Walker; Anath C. Lionel; Peter Pasceri; Daniele Merico; Ryan K.C. Yuen; Karun K. Singh; James Ellis; Stephen W. Scherer
• Keywords: Candidate gene

Autism spectrum disorder (ASD) is phenotypically and genetically heterogeneous. We present a CRISPR gene editing strategy to insert protein tag 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 iPSCs allowed production excitatory neurons, mutant proteins were not detectable. RNA sequencing revealed convergence several neuronal networks. Using both patch-clamp multi-electrode array approaches, the electrophysiological deficits measured distinct different mutations. However, they culminated in consistent reduction synaptic activity, including reduced spontaneous postsynaptic current frequencies AFF2/FMR2-, ASTN2-, ATRX-, KCNQ2-, SCN2A-null neurons. Despite ASD susceptibility belonging ontologies, isogenic resources can reveal common phenotypes, such as connectivity.