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ARS Home » Midwest Area » Ames, Iowa » National Animal Disease Center » Infectious Bacterial Diseases Research » Research » Research Project #442006

Research Project: Isolation and CRISPRI-based Functional Genomics Approaches for Pathogenic Leptospira Isolated From Environmental Samples

Location: Infectious Bacterial Diseases Research

Project Number: 5030-32000-233-014-N
Project Type: Non-Funded Cooperative Agreement

Start Date: Aug 1, 2022
End Date: May 31, 2023

Objective:
Obtain pathogenic leptospires from complex environmental samples (soil and biofilm), through standardization and amelioration of specific protocols, and identify genes important for biofilm formation.

Approach:
1. Soil sample collection: Soil samples previously collected by ARS from an endemic site for leptospirosis will be analyzed according to protocols previously published by ARS. 2. Search for candidate genes for biofilm formation: In silico analysis to identify candidate genes from Leptospira that play a role in biofilm formation. CRISPRi assay. 3. As per ARS previously published protocols, CRISPR interference will be used for gene silencing of selected targets in pathogenic Leptospira. The characterization of the mutants will be done by biofilm formation, confocal microscopy, electron microscopy, fluorescent antibody test, and immunoblot.