Fig. 3: Phenotype proportions in knockout models of cardiac candidate genes.

A Validation workflow encompassing zygotic microinjections using a HdrR (myl7::eGFP; myl7::H2A-mCherry) reporter line, followed by phenotypic classification of embryos with normal developed hearts and embryos with cardiac-specific phenotypes 4 days post-fertilization (dpf). B Proportion (bars) and counts (values) of cardiac-affected and normally developed embryos after CRISPR-Cas9-mediated knockout of indicated candidate genes versus control (mock injection). Phenotypic proportions of crispants were determined from 54 to 100 embryos and compared to 303 mock-injected control embryos. The mean and standard deviation of the control group were calculated based on seven technical replicates. C Independent replication using base editing: phenotypic distribution (proportion/bars and counts/values) resulting from targeted gene editing mediated by introducing premature termination codons via the cytosine base editor evoBE4max and a set of distinct guide RNAs targeting the same genes as in (B). Phenotypic proportions of editants were determined from 50 to 75 embryos and compared to 105 mock-injected control embryos. The mean and standard deviation of the control group were calculated based on three technical replicates.