| 论文作者 |
Wang, Y; Qi, T; Liu, JT; Yang, Y; Wang, ZW; Wang, Y; Wang, TY; Li, MM; Li, MQ; Lu, DR; Chang, ACY; Yang, L; Gao, S; Wang, YM; Lan, F |
| 摘要 |
The CRISPR-Cas system can treat autosomal dominant diseases by nonhomologous end joining (NHEJ) gene disruption of mutant alleles. However, many single-nucleotide mutations cannot be discriminated from wild -type alleles by current CRISPR-Cas systems. Here, we functionally screened six Cas12j nucleases and determined Cas12j-8 as an ideal genome editor with a hypercompact size. Cas12j-8 displayed comparable activity to AsCas12a and Un1Cas12f1. Cas12j-8 is a highly specific nuclease sensitive to single-nucleotide mismatches in the protospacer adjacent motif (PAM)-proximal region. We experimentally proved that Cas12j-8 enabled allele -specific disruption of genes with a single-nucleotide polymorphism (SNP). Cas12j-8 recognizes a simple TTN PAM that provides for high target site density. In silico analysis reveals that Cas12j-8 enables allele-specific dis-ruption of 25,931 clinically relevant variants in the ClinVar database, and 485,130,147 SNPs in the dbSNP data-base. Therefore, Cas12j-8 would be particularly suitable for therapeutic applications. |
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