Fig. 3.

miR-125a1^−/− promote vessel sprouting.

(A) Schematic representation of the generation of the miR-125a1^−/− CRISPR/Cas9 mutants. The 5′ and 3′ regions of the zebrafish miR-125a1 gene contain protospacer adjacent motifs (PAM; Green). Guide strand RNA (red) were generated against target oligonucleotides (black) down-stream of these PAM sites. TracrRNA (blue) enables the homing of the Cas9 protein to the targeted PAM site. (B) Genotyping of miR-125a1^+/+, miR-125a1^+/− and miR-125a1^−/− embryos, respectively (Table S2 primer sequences). (C) The frequency of genotype variants; miR-125a1^+/+, miR-125a1^+/− and miR-125a1^−/− embryos, shown as a percentage of total number of in-crossed miR-125a1^+/− progeny (n = 57). (D) Representative image of miR-125a1^+/+ and miR125a1^−/− embryos, showing aberrant ISV sprouting (red arrowheads) in miR-125a1^−/− embryos. (E) The frequency of intersegmental vessel hyper-branching. (F) The incidence of no, mild, moderate and severe vascular hyper-branching caused by loss of miR-125a1. Data are represented as phenotype frequencies. *** p < .005 significantly different to control. See also Fig. S5. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)