Altered neurogenesis and disrupted expression of synaptic proteins in prefrontal cortex of SHANK3-deficient non-human primate

We used the Cas9/sgRNA method to disrupt SHANK3 gene in cynomolgus monkeys (Macaca fascicularis). SHANK3 has an array of mRNA isoforms due to multiple promoters and extensive alternative splicing of coding exons. We therefore simultaneously targeted two sites of SHANK3 to induce a large deletion that could completely disrupt all SHANK3 isoforms. All three offspring were positive for mutations at site 2 of exon 12. We aimed to analyze the ratio of indels in different tissues. Only small indels can be estimated by deep amplicon sequencing. PCR products were added with barcodes in both ends and pooled for each animal. The samples were purified and used for Illumina sequencing at Novogene. The sequences from three animals were uploaded.