Products/Services Used | Details | Operation |
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Gene Services> | The full-length zebrafish codon-optimized bpNLS-ScCas9-bpNLS sequence was synthesized by GenScript, AllcrRNAsandtracrRNAswerechemicallysynthesized by GenePharma, Shanghai, China or GenScript, Nanjing, China Concentrated protein was confirmed by western blotting with His-tag antibody (GenScript, Nanjing, China) and stored at−80◦C. | Get A Quote |
The CRISPR/Cas9 system has been widely used for gene editing in zebrafish. However, the required NGG protospacer adjacent motif (PAM) of Streptococcus pyogenes Cas9 (SpCas9) notably restricts the editable range of the zebrafish genome. Recently, Cas9 from S. canis (ScCas9), which has a more relaxed 50-NNG-30 PAM, was reported to have activities in human cells and plants. However, the editing ability of ScCas9 has not been tested in zebrafish. Here we characterized and optimized the activity of ScCas9 in zebrafish. Delivered as a ribonucleoprotein complex, ScCas9 can induce mutations in zebrafish. Using the synthetic modified crRNA:tracrRNA duplex instead of in vitro-transcribed single guide RNA, the low ... More