Fig. 2.

Efficient targeted mutagenesis of brachyury ortholog. (A,B) Whereas animals injected with nls-EGFP mRNA display no phenotype at 21 days post-fertilization (A), their siblings injected with sgRNA directed against a putative brachyury ortholog and cas9 display a dramatic shortening along the anteroposterior axis (B). (C) This phenotype persists at 7 months post-fertilization, as an sgRNA-injected animal (bottom) shows a severe reduction in length of the posterior body compared with an nls-EGFP-injected sibling (top). (D) Sequencing of the targeted region from two brachyury RGN-injected 21-day-old whole larvae displaying a severe phenotype as in B reveal a high indel frequency at the targeted site. (E) Sequencing of the targeted region from DNA extracted from fin clips of two brachyury RGN-injected 5-month-old animals displaying a severe phenotype as in C reveals a high indel frequency at this site.