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JoVE Journal Genetics
Anvendelse af CRISPR-interferens (CRISPRi) til genhæmning i patogene arter af Leptospira
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Anvendelse af CRISPR-interferens (CRISPRi) til genhæmning i patogene arter af 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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Her beskrives anvendelsen af CRISPR-interferens (CRISPRi) til specifik genhæmning i Leptospira-arter. Leptospira celler omdannes ved bøjning med plasmider udtrykke dCas9 (katalytisk "døde" Cas9) og en enkelt-guide RNA (sgRNA), der er ansvarlig for base parring til det ønskede genomiske mål. Metoder til validering af genhæmning præsenteres.

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Genetik Leptospira leptospirose mutagenesis CRISPR interferens genhæmning bøjning
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