Modification of LbCas12a enzyme for improving thermal stability and application of LbCas12a enzyme

Through genetic modification and mutant screening, thermally stable LbCas12a enzyme was obtained, which solved the problem of enzyme inactivation under high temperature conditions, improved the reaction efficiency and specificity, and expanded the scope of application.

CN119709699AActive Publication Date: 2025-03-28CHINESE ACAD OF INSPECTION & QUARANTINE
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Patent Information

Application Number
CN202411956609.6
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2024-12-29
Publication Date
2025-03-28
Estimated Expiration
2044-12-29

AI Technical Summary

Technical Problem

The inactivation of LbCas12a enzyme under high temperature conditions affects the reaction efficiency and specificity of the CRISPR/Cas12a system, resulting in low detection specificity and risk of contamination.

Method used

Through genetic modification and mutant screening, a thermally stable LbCas12a artificial engineered enzyme, called CAIQ-Cas12a enzyme, was obtained. This enzyme has good activity at 54°C and its thermal stability has been improved by about 8°C.

Benefits of technology

It improves the thermal stability of LbCas12a enzyme, enhances its reaction ability in CRISPR/Cas12a system, reduces the risk of inactivation, and expands its application range and use effect.

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Abstract

The invention relates to an LbCas12a artificially modified enzyme for improving thermal stability. The amino acid sequence of the LbCas12a artificially modified enzyme is as shown in SEQ ID NO.1, and the nucleotide gene sequence for coding the LbCas12a artificially modified enzyme is as shown in SEQ ID NO.2. The invention also relates to a preparation method of the LbCas12a artificially modified enzyme. Compared with an original LbCas12a enzyme, the mutant Y554C obtained through screening has the advantages that the thermal stability is improved by about 8 DEG C, the reaction requirement in a CRISPR / Cas12a system can be met, the application range of the LbCas12a enzyme is widened, the use effect of the LbCas12a enzyme is improved, and the mutant Y554C can be used for commercial application.
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