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JoVE Journal Genetics
Aplicación de interferencia CRISPR (CRISPRi) para silenciamiento de genes en especies patógenas de Leptospira
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Aplicación de interferencia CRISPR (CRISPRi) para silenciamiento de genes en especies patógenas de 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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Aquí, se describe la aplicación de la interferencia CRISPR (CRISPRi) para el silenciamiento de genes específicos en especies de Leptospira. Las células de leptospira se transforman por conjugación con plásmidos que expresan dCas9 (catalíticamente "muerto" Cas9) y un ARN de guía única (sgRNA), responsable del emparejamiento de bases a la diana genómica deseada. Se presentan métodos para validar el silenciamiento génico.

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Genética Número 174 Leptospira,leptospirosis mutagénesis interferencia CRISPR silenciamiento génico conjugación
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