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Genetics
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JoVE Journal Genetics
Application of CRISPR Interference (CRISPRi) for Gene Silencing in Pathogenic Species of Leptospira
 

Application of CRISPR Interference (CRISPRi) for Gene Silencing in Pathogenic Species of Leptospira

Article DOI: 10.3791/62631-v • 14:49 min • August 14th, 2021
August 14th, 2021 •
1,2, 1, *2, *1
1Infectious Bacterial Diseases Research Unit, National Animal Disease Center, Agricultural Research Service, United States Department of Agriculture, 2Laboratório de Desenvolvimento de Vacinas, Instituto Butantan
* These authors contributed equally

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Here, the application of CRISPR interference (CRISPRi) for specific gene silencing in Leptospira species is described. Leptospira cells are transformed by conjugation with plasmids expressing dCas9 (catalytically "dead" Cas9) and a single-guide RNA (sgRNA), responsible for base pairing to the desired genomic target. Methods to validate gene silencing are presented.

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CRISPR Interference CRISPRi Gene Silencing Pathogenic Species Leptospira Leptospirosis Neglected Disease Bacteria Infection Research Team National Animal Disease Center Ames Iowa Pathogenic Mechanisms Specific Genes Expression Growth Media Fastidious Leptospiras Rick Hornsby Propagation Media Louis Fernandez Target Genes Silencing
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