Construction of high-fidelity CRISPR/ASCPF1 mutant and uses thereof

Abstract

A high-fidelity AsCpf1 mutant is obtained by performing a mutation on arginine at position 951 and/or 955 of a AsCpf1 protein amino acid sequence and replacing the same with an amino acid free of forming a hydrogen bond with DNA of a target site; and the amino acid sequence thereof is shown in SEQ ID NOS: 1-3. The encoding gene of the AsCpf1 mutant has a nucleotide sequence as shown in SEQ ID NO: 4 and can be used in the construction of a CRISPR/AsCpf1 gene editing system. A CRISPR/AsCpf1 gene editing system includes a gene encoding a AsCpf1 protein, and the AsCpf1 protein is the AsCpf1 mutant mentioned above. The CRISPR/AsCpf1 gene editing system can be used in lowering an off-target effect of gene editing. The novel AsCpf1 mutant not only retains the gene editing efficiency of wild-type AsCpf1, but also has a higher specificity than the wild-type AsCpf1.

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