Xenbase Image ID: 152312

Figure 7. Example of target gene analysis according to our workflow. We designed an sgRNA targeting activin receptor-like kinase 2 (alk2). (A) Representative phenotypes produced by Cas9 mRNA and sgRNA (upper panel). Most phenotypes exhibited a partial loss of ventral fin tissue accompanied by the hypertrophic anus. Normal means no alteration in the ventral fin or anus. The total numbers of individuals are shown at the top of each graph. In vitro fertilization (IVF) indicates control embryos without injection. Frequencies of alk2-disrupted phenotypes from three independent experiments are shown in under panel. (B) RGEN-RFLP analysis of somatic mutation rates. (C) Sanger DNA sequencing of somatic mutation alleles from five phenotypes. In total, 107 clones were sequenced. (D) Off-target analysis using heteroduplex mobility assay (HMA). Wt, wild type; P, phenotype. Image published in: Shigeta M et al. (2016) Copyright © 2016. Image reproduced with permission of the Publisher, John Wiley & Sons. Experiment + Assay Source Phenotypes and Disease Xtr Wt + acvr1 CRISPR + NF41 (morphology) fig.7.a Anatomical Phenotype abnormally ventrally curved tail increased size of the proctodeum Larger Image Printer Friendly View

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