<p>Autism Spectrum Disorder 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 10 ASD-relevant genes (<a href="https://www.ncbi.nlm.nih.gov/gene/?term=2334">AFF2/FMR2</a>, <a href="https://www.ncbi.nlm.nih.gov/gene/?term=3730">ANOS1</a>, <a href="https://www.ncbi.nlm.nih.gov/gene/?term=23245">ASTN2</a>, <a href="https://www.ncbi.nlm.nih.gov/gene/?term=546">ATRX</a>, <a href="https://www.ncbi.nlm.nih.gov/gene/?term=775">CACNA1C</a>, <a href="https://www.ncbi.nlm.nih.gov/gene/?term=57680">CHD8</a>, <a href="https://www.ncbi.nlm.nih.gov/gene/?term=9228">DLGAP2</a>, <a href="https://www.ncbi.nlm.nih.gov/gene/?term=3785">KCNQ2</a>, <a href="https://www.ncbi.nlm.nih.gov/gene/?term=6326">SCN2A</a>, <a href="https://www.ncbi.nlm.nih.gov/gene/?term=10178">TENM1</a>). Neurogenin 2 (<a href="https://www.ncbi.nlm.nih.gov/gene/?term=63973">NGN2</a>)-directed induction of iPSCs allowed production of 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.</p>

CRISPR-Mediated ASD Gene Knockout Reduces Neuronal Activity

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