Disruption of src and tks5a genes by CRISPR/Cas9 affects the PLLp migration. Two sgRNAs targeting two different exons (25ng/µl) within each gene were co-injected with Cas9 mRNA (300ng/µl) into cldnb:EGFP embryos and the embryos raised. Injected fish were inbred and at 48hpf WT embryos show the normal pattern of the migrating primordium and neuromasts deposition (A), whereas src CRISPR (B), tks5a CRISPR (C) and tks5a/ (D) embryos show a disruption of the primordium migratory ability and PLL morphogenesis. Arrows indicate the position of the primordium in tks5a/ embryos; asterisk indicates the position of the last deposited neuromast. (E) Sequence confirmation of tks5a/alleles. Allele 1 has an 8-bp insertion and a 1-bp deletion, which changed amino acid valine at position 171 to threonine and truncates the protein after 27 amino acids. Allele 2 has a 21-bp insertion and a 4-bp deletion, which changed amino acid proline at position 369 to glutamine and truncated the protein after 14 amino acids.
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