Casdelta variant with improved editing efficiency and application thereof

By mutating specific amino acid positions of the Cas protein, a Cas protein mutant with enhanced activity was developed, overcoming the shortcomings of the CRISPR/Cas system in terms of PAM range, molecular size, and multiplexing efficiency, and achieving more efficient gene editing results.

CN121087015APending Publication Date: 2025-12-09CHINA AGRI UNIV
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Patent Information

Application Number
CN202511594006.0
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-11-03
Publication Date
2025-12-09

AI Technical Summary

Technical Problem

Existing CRISPR/Cas systems have room for improvement in terms of PAM range, molecular size, RNA requirements, and multiplex editing efficiency, and there is a need to improve the editing activity and stability of Casδ protein.

Method used

By modifying amino acids on the basis of wild-type Cas protein, we developed Cas protein mutants with enhanced activity. These mutants include modifications at specific amino acid positions to enhance catalytic activity, binding activity, specificity, and stability, and can combine with other molecules to form fusion proteins.

Benefits of technology

It improves the target sequence cleavage and editing activity of Cas protein, enhances its binding ability with gRNA, reduces off-target risk, and achieves more efficient gene editing.

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Abstract

The invention relates to the field of nucleic acid editing, in particular to the technical field of regularly clustered interval short palindromic repeat (CRISPR). In particular, the invention relates to a mutant of a Cas effector protein, a fusion protein comprising the mutant, and nucleic acid molecules encoding the same. The invention also relates to complexes and compositions for nucleic acid editing (e.g., gene or genome editing). The invention also relates to methods for nucleic acid editing (e.g., gene or genome editing) using the proteins or fusion proteins comprising the invention. Compared with a wild type Cas protein, the Cas protein mutant disclosed by the invention has better activity, such as higher cleavage activity, stronger target site recognition capability and higher target sequence editing activity.
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