CRISPR screening of autism risk genes in brain organoids
Organoids were generated from cells infected with pooled lentivirus library. The lentiviral libraries deliver dual-gRNAs targeting 36 high-risk ASD genes in parallel. With or without 4-OHT induction of eCas9 expression, organoids were dissociated at specific developmental stages. Single cells were sorted and bulk genomic DNA was extracted. Target amplification was performed using genomic DNA to analyze gRNA and unique clone barcode representation.