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

4,613 Views

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14:49 min

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August 14, 2021

DOI:

10.3791/62631-v

DOI:
10.3791/62631-v

•

14:49 min
August 14, 2021

•

7 Views
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1Infectious Bacterial Diseases Research Unit, National Animal Disease Center, Agricultural Research Service,United States Department of Agriculture, 2Laboratório de Desenvolvimento de Vacinas,Instituto Butantan

Summary

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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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Cite this Article

Fernandes, L. G. V., Hornsby, R. L., Nascimento, A., Nally, J. E. Application of CRISPR Interference (CRISPRi) for Gene Silencing in Pathogenic Species of Leptospira. J. Vis. Exp. (174), e62631, doi:10.3791/62631 (2021).

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