Low-immunogenicity LwCas13a protein mutant and application thereof

By performing Q436C, K560D and K76C mutations on the LwCas13a protein, the immunogenicity problem of the Cas13 protein in cattle was solved, and the stable expression and efficient gene editing of the low-immunogenic LwCas13a protein in cattle were achieved.

CN120738153AActive Publication Date: 2025-10-03INNER MONGOLIA UNIVERSITY
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Patent Information

Application Number
CN202511269339.6
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-09-08
Publication Date
2025-10-03
Estimated Expiration
2045-09-08

AI Technical Summary

Technical Problem

Long-term expression of Cas13 protein in large mammals has immunogenicity issues and may be recognized as a target of attack by the immune system, leading to instability of immune response and targeted RNA regulation.

Method used

A low-immunogenic LwCas13a protein mutant was designed by introducing mutations such as Q436C, K560D, and K76C into the wild-type LwCas13a protein to reduce its binding affinity to bovine MHC-I class molecules while maintaining the structural stability and nuclease activity of the protein.

Benefits of technology

The immunogenicity of LwCas13a protein in cattle was significantly reduced, ensuring the long-term stable expression of the protein and the regulatory ability of targeted RNA, maintaining the gene editing efficiency, and improving the safety and effectiveness of the application.

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Abstract

The invention relates to the technical field of gene engineering, in particular to a low-immunogenicity LwCas13a protein mutant and application thereof. According to the LwCas13a protein mutant disclosed by the invention, one or more mutations of Q436C, K560D and K76C are generated on a wild type LwCas13a protein. According to the LwCas13a protein mutant disclosed by the invention, the immunogenicity of the LwCas13a protein in a cattle body is remarkably reduced, so that the recognition and reaction of an immune system are reduced; the LwCas13a protein mutant disclosed by the invention not only does not influence the activity of LwCas13a nuclease, but also can maintain the effective expression and function of the LwCas13a nuclease in bovine cells; the gene editing efficiency of the LwCas13a protein mutant provided by the invention is equivalent to that of a wild type, and the interference efficiency is not obviously influenced.
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