Figure 6.
AHR2b CRISPR-Cas9 off-target analysis. A. Nucleotide sequence alignment of the AHR2b CRISPR-Cas9 target sequence with the corresponding regions of closely related genes. AHR1a, AHR2a, AHR1b, and AHRR have 4, 4, 6, and 3 mismatches to AHR2b in this region, respectively. B. Surveyor nuclease detection of mutations in AHR1a, AHR2a, AHR1b, and AHRR in the AHR2b CRISPR-Cas9 target region. Digested fragments are indicated by a *. Each lane represents a pool of 5 embryos from which genomic DNA fragments were amplified. U: uninjected control, 1–4: CRISPR-Cas9 injected embryos. AHR1a, AHR2a, and AHRR PCR products from uninjected and injected embryos were sequenced.