Skip to main content
. Author manuscript; available in PMC: 2022 Mar 1.
Published in final edited form as: Dev Biol. 2020 Dec 5;471:1–9. doi: 10.1016/j.ydbio.2020.12.001

Fig. 4. Hoxb5b sgRNA/Cas9 mediated somatic mutagenesis phenocopies the bifurcated foregut defects found in Hoxb5b morphants.

Fig. 4.

(A) Forward and reverse primer sequences are shown (underlined). Hoxb5b sgRNA1 (blue) and sgRNA2 (green) genomic target sequences are shown. Protospacer adjacent motif-PAM sequence (pink). (B) T7 endonuclease I (T7EI) assay. (Lane 2) marker. (Lane 1) untreated and (lane 3,10) T7EI treated amplicons from control embryos. (Lane 5–9) amplicons from embryos injected with hoxb5b sgRNA1/Cas9 or (Lane 4,11–13) hoxb5b sgRNA2/Cas9 were digested by T7EI. Live images of (C) Control, and (D, E) sgRNA1/Cas9 and sgRNA2/Cas9 injected Tg(sox17:EGFP) at 30 hpf. Somatic disruption of the Hoxb5b locus by two separate sgRNAs phenocopies foregut defects in Hoxb5b morphants. Anterior to the top, results are from 2 independent experiments.