Fig. 3
Targeting ppp2r3b in zebrafish using gene-editing. (A) ppp2r3b gene structure showing the location of primers used for genotyping (red arrows) and sgRNA (green) used for gene-editing. Below, examples of mutant sequence reads cloned from pooled F0 embryos. The sgRNA site is highlighted in yellow within the wild-type sequence. Inserted and deleted nucleotides are highlighted in pink and blue, respectively. (B) Sequence chromatograms showing the homozygous wild-type reference and alternative alleles, and the homozygous and heterozygous mutant reads. (C, D) RT-PCR showing expression of ppp2r3b using primers located within (C, D) exons 1?3 (band at expected size, 491 bp) or (D) or exons 1?7 (band at expected size, 1000 bp) at the indicated stages. |