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

Deneault, E.; White, S. H.; Rodrigues, D. C.; Ross, J.; Faheem, M.; Zaslavsky, K.; Wang, Z.; Alexandrova, R.; Pellecchia, G.; Wei, W.; Piekna, A.; Kaur, G.; Howe, J. L.; Kwan, V.; Thiruvahindrapuram, B.; Walker, S.; Pasceri, P.; Merico, D.; Yuen, R. K.; Singh, K. K.; Ellis, J.; Scherer, S. W.

doi:10.1101/344234

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bioRxiv · 10.1101/344234

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

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Abstract

Autism 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.

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Deneault, E., White, S. H., Rodrigues, D. C., Ross, J., Faheem, M., Zaslavsky, K., Wang, Z., Alexandrova, R., Pellecchia, G., Wei, W., Piekna, A., Kaur, G., Howe, J. L., Kwan, V., Thiruvahindrapuram, B., Walker, S., Pasceri, P., Merico, D., Yuen, R. K., Singh, K. K., Ellis, J., Scherer, S. W.. 2018-06-11. Complete Disruption of Autism-Susceptibility Genes by Gene-Editing Predominantly Reduces Functional Connectivity of Isogenic Human Neurons. https://doi.org/10.1101/344234

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