Guided genetic screen to identify genes essential in the regeneration of hair cells and other tissues

The screening pipelines. a The pipeline for screening mutations obtained by retroviral integration. Homozygous embryos from the F2 generation were used for screening. Mutations obtained by ENU or gene/protein trap were also screened using homozygotes from the F2 generation. b The pipeline for screening mutations obtained by CRISPR/Cas9. Multiplex targeting was usually used to generate CRISPR/Cas9 mutations, by co-injecting 2–6 single-guide RNA (sgRNA) together with Cas9 mRNA. Compound heterozygotes in the F1 generation were used for screening when the sgRNAs had sufficient measured somatic activity. Homozygotes or compound heterozygotes from the F2 generation were used when there were not enough compound heterozygotes found in the F1 generatio