Figure 1

Quantification of phenotypes and gRNA efficacy. (A) Amplicons for each gRNA target were generated by PCR and then TOPO-cloned to generate a library of targeted genomic DNA sequences. (B) Individual clones were amplified by colony PCR and sent for sequencing. (C) Sequences were aligned to wt to determine the incidence of indel mutations. (D) For most targets, we are unable to isolate wt DNA by cloning, indicating a very high degree of mutational efficiency (with the exception of emilin1b gRNA 2, which appears to be non-functional). (E) Quantification of cardiomegaly phenotypes for all genes at 2 and 5 dpf. (F) Quantification of axial curvature defects for all genes at 2 and 5 dpf. (G) Quantification of hemorrhage defects for all genes at 2 and 5 dpf.