A Highly Specific CRISPR-Cas12j Nuclease Enables Allele-specific Genome Editing

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2023 Feb 10

PMID 36763662

Authors

Yao Wang

Tao Qi

Jingtong Liu

Yuan Yang

Ziwen Wang

Ying Wang

Tianyi Wang

Miaomiao Li

MingQing Li

Daru Lu

Alex Chia Yu Chang

Li Yang

Song Gao

Yongming Wang

Feng Lan

Affiliations

Soon will be listed here.

Abstract

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 disruption of 25,931 clinically relevant variants in the ClinVar database, and 485,130,147 SNPs in the dbSNP database. Therefore, Cas12j-8 would be particularly suitable for therapeutic applications.

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