Extended Data Fig. 8: Depletion of PTC-bearing mRNA impairs GCR in capn3a^−/− embryos.

a, b, WISH was performed to assess the liver sizes in 3.5-dpf capn3a^−/− embryos injected with different reagents as indicated. c, Relative expression of capn8 and capn12 in wild-type embryos injected with different reagents. d–f, qRT–PCR (d), western blot analysis (e) or WISH (f) in wild-type embryos injected with different reagents. Actb1 was used as a loading control. g, Top, a capn3a mutant mRNA (capn3a^3pm), which is resistant to the degradation mediated by Cas13a gRNA, was designed. Red letters denote nucleotide mutations in the target sites; red stars denote the target sites of three gRNAs. Bottom, statistical analysis of the average liver sizes in wild-type embryos injected with or without Cas13a and non-specific gRNA (NS), Cas13a and gRNA (G), or Cas13a and gRNA plus capn3a^3pm mRNA at 3.5 dpf. h, Statistical analysis of the average liver sizes in capn3a^−/− embryos injected with or without Cas13a and non-specific gRNA (NS) or Cas13a and gRNA (G) at 3.5 dpf. Each experiment was repeated three times with similar results, and a representative result is shown. n indicates the number of zebrafish embryos in each group. Data are mean ± s.d. from three biological replicates. *P < 0.05, **P < 0.01, ***P < 0.001, two-tailed t-test.