CRISPR-Cas13a-based crRNA sequence and plasmid for targeting vibrio parahaemolyticus and application of crRNA sequence and plasmid

By designing crRNA targeting Vibrio parahemolyticus and constructing CRISPR-Cas13a bactericidal plasmid, the cleavage activity of the Cas13a protein was used to solve the prevention and control problems of Vibrio parahemolyticus, achieving efficient and specific bactericidal effects, and avoiding drug resistance.

CN120442629APending Publication Date: 2025-08-08GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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Patent Information

Application Number
CN202510595144.4
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-05-09
Publication Date
2025-08-08

AI Technical Summary

Technical Problem

The prior art is difficult to efficiently and specifically prevent and control Vibrio parahaemolytic. The traditional methods have problems with unstable bactericidal effects and drug resistance, and the application of the CRISPR-Cas system in Gram-negative bacteria is not yet mature.

Method used

CrRNA specifically targeting the T2RHS-Nuc gene of Vibrio parahaemolytica was designed, and a bactericidal plasmid was constructed for the CRISPR-Cas13a system was introduced into Vibrio parahaemolytica through E. coli, and the cleavage activity of the Cas13a protein was used to target the killing strain.

Benefits of technology

The specific killing of Vibrio parahaemolyticus was achieved, the drug resistance problem was avoided, and a safe and efficient prevention and control plan was provided.

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Abstract

The invention discloses a crRNA sequence and a plasmid for targeting vibrio parahaemolyticus based on CRISPR-Cas13a (clustered regularly interspaced short palindromic repeats) and application of the crRNA sequence and the plasmid. According to the invention, crRNA of a targeted T2RHS-Nuc gene is designed, a plasmid of tetracycline-induced expression Cas13a protein is established, the plasmid is introduced into vibrio parahaemolyticus under the assistance of escherichia coli and targets a target gene, and after mRNA of the target gene is recognized, targeted cleavage activity is generated, so that the affiliated cleavage activity of Cas13a is activated, a transcript of a bacterial strain is randomly cleaved in the bacterial body, and the expression of the Cas13a protein is promoted. When the transcript is necessary for the growth of the vibrio parahaemolyticus, the specific death of the vibrio parahaemolyticus can be caused. The crRNA and Cas13a protein expression vector and the sterilization method thereof provided by the invention do not induce vibrio parahaemolyticus to generate drug resistance, are a good alternative method for vibrio parahaemolyticus disease source control by using a traditional bactericide, and have a very good application prospect.
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