Marker-less deletion of cctA gene of Clostridium chauvoei

Sanjay Kumar Gupta; Sameer Dixit; Saroj K Dangi; Gurpreet Kaur; Mohmad Mashooq; Kumaragurubaran Karthik; Mihir Sarkar; Sonalika Mahajan; Viswas Konasagara Nagaleekar

doi:10.1016/j.anaerobe.2019.102116

Marker-less deletion of cctA gene of Clostridium chauvoei

Anaerobe. 2020 Feb:61:102116. doi: 10.1016/j.anaerobe.2019.102116. Epub 2019 Nov 8.

Authors

Sanjay Kumar Gupta¹, Sameer Dixit¹, Saroj K Dangi¹, Gurpreet Kaur¹, Mohmad Mashooq¹, Kumaragurubaran Karthik¹, Mihir Sarkar², Sonalika Mahajan³, Viswas Konasagara Nagaleekar⁴

Affiliations

¹ Division of Bacteriology and Mycology, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122, India.
² Division of Physiology and Climatology, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122, India.
³ Division of Veterinary Biotechnology, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122, India.
⁴ Division of Bacteriology and Mycology, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly, 243122, India. Electronic address: kn.viswas@icar.gov.in.

PMID: 31711886
DOI: 10.1016/j.anaerobe.2019.102116

Abstract

Clostridium chauvoei causes blackleg disease in domestic animals, especially cattle and sheep. The pathogen produces several toxins including CctA - a hemolysin and protective antigen. Molecular pathogenesis of the disease is poorly understood, possibly due to lack of genetic manipulation tools for C. chauvoei. In the present study, we report the marker-less deletion of cctA gene using the CRISPR-Cas9 system. The C. chauvoei cctA deletion mutant had negligible hemolytic and significantly reduced cytotoxic activities. To the best of our knowledge, this is the first report of genetic manipulation of C. chauvoei. The method we used in this study can be applied for genetic manipulation of C. chauvoei to better understand the pathogenesis and genetics of the pathogen.

Keywords: CRISPR-Cas9; Clostridium chauvoei; Genome editing; cctA.

MeSH terms

Animal Diseases / microbiology*
Animal Diseases / prevention & control
Animals
Animals, Domestic
Anti-Bacterial Agents / pharmacology
Antibiotic Prophylaxis
Bacterial Proteins / genetics*
CRISPR-Cas Systems
Clostridium Infections / veterinary*
Clostridium chauvoei / drug effects
Clostridium chauvoei / genetics*
Gene Deletion*
Gene Editing
Hemolysin Proteins / genetics*
Hemolysis
Mutation

Substances

Anti-Bacterial Agents
Bacterial Proteins
Hemolysin Proteins