HIGH-FIDELITY CAS9 VARIANTS AND APPLICATIONS THEREOF
收藏国家林业和草原科学数据中心2023-02-12 更新2024-03-07 收录
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To address the limitations deriving from the unspecific genomic cleavages of the Streptococcus pyogenes Cas9 (SpCas9) and to identify variants with higher cleavage fidelity， the present invention describes a yeast-based assay which allows to simultaneously evaluate the on- and off-target activity towards two engineered genomic targets. The screening of SpCas9 variants obtained by random mutagenesis of the Red-II domain allowed the identification of hits with increased on/off ratios. The best performing nuclease， evoCas9， was isolated through the combination of the identified mutations within a single variant. Side by side analyses with previously reported rationally designed variants demonstrated a significant improvement in fidelity of evoCas9 of the present invention.
提供机构：
国家林业和草原科学数据中心
创建时间：
2023-02-12
