SRA

Here, we optimize and characterize SpG and SpRY systems in zebrafish. Using purified protein combined with synthetic modified gRNA, nearly PAM-less sites could be targeted efficiently in zebrafish genome, which suggested that SpG and SpRY can be efficiently used in vertebrate. In addition, we compared SpCas9, SpG, SpRY, VQR and ScCas9 editing activity bearing different PAMs and gave an advice for selection of CRISPR-Cas toolbox to precise edit genome in zebrafish. Altogether, our results significantly improved the CRISPR-Cas targeting genomic landscape in zebrafish model and switch on the application of SpG and SpRY toolbox in other in vivo systems.