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Journal / Genetics / Anwendung der CRISPR-Interferenz (CRISPRi) zur…
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JoVE Journal Genetics
Anwendung der CRISPR-Interferenz (CRISPRi) zur Gen-Silencing bei pathogenen Leptospira-Spezies
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Anwendung der CRISPR-Interferenz (CRISPRi) zur Gen-Silencing bei pathogenen Leptospira-Spezies

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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Hier wird die Anwendung der CRISPR-Interferenz (CRISPRi) für spezifisches Gen-Silencing bei Leptospira-Spezies beschrieben. Leptospira-Zellen werden durch Konjugation mit Plasmiden transformiert, die dCas9 (katalytisch "totes" Cas9) und eine Single-Guide-RNA (sgRNA) exprimieren, die für die Basenpaarung zum gewünschten genomischen Ziel verantwortlich ist. Methoden zur Validierung des Gen-Silencing werden vorgestellt.

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Genetik Ausgabe 174 Leptospira Leptospirose Mutagenese CRISPR-Interferenz Gen-Silencing Konjugation
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