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Updated: Jul 7, 2025

Application of CRISPR Interference CRISPRi for Gene Silencing in Pathogenic Species of Leptospira
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A robust CRISPR interference gene repression system in Vibrio parahaemolyticus.

Taoyuan Jiang1, Yuhuan Li2, Wencong Hong3

  • 1Department of Respiratory Medicine, Nan'an Hospital, 330, Ximei Residential District, Xinhua Street, Quanzhou, Fujian Province, China. jiangty01@163.com.

Archives of Microbiology
|December 26, 2023
PubMed
Summary

Researchers developed a new CRISPR interference (CRISPRi) system for Vibrio parahaemolyticus, enabling scalable and tunable gene repression. This tool aids in studying the significant seafood-associated pathogen.

Keywords:
CRISPRiEssential geneGene repressionVibrio parahaemolyticus

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Genetics

Background:

  • Vibrio parahaemolyticus causes widespread seafood-associated gastroenteritis and aquaculture losses.
  • Limited protein depletion strategies hinder research into V. parahaemolyticus.

Purpose of the Study:

  • To develop a novel CRISPR interference (CRISPRi) system for targeted gene repression in V. parahaemolyticus.
  • To establish a scalable, tunable, and reversible gene repression method for this important pathogen.

Main Methods:

  • Construction of a CRISPRi system using the pVv3 shuttle vector and a nuclease-null Cas9 (dCas9) from Streptococcus pyogenes.
  • Induction of gene repression using IPTG and arabinose, with scalability demonstrated using multiple single-guide RNAs (sgRNAs).
  • Tunability achieved by adjusting inducer concentrations and reversibility confirmed by inducer removal.

Main Results:

  • Successful establishment of an inducible CRISPRi system in V. parahaemolyticus.
  • Demonstrated scalability of gene repression using multiple sgRNAs.
  • Precise control over repression levels and confirmed reversibility of the system.

Conclusions:

  • The developed CRISPRi system offers a simple and effective method for genome-wide gene repression in V. parahaemolyticus.
  • This tool will significantly advance research on V. parahaemolyticus, including its physiology, pathogenesis, and drug target identification.