Fig. 1.

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 generation