Catalytically Active Cas9 Mediates Transcriptional Interference to Facilitate Bacterial Virulence
| Autor: | Jessie E. Wozniak, Andrés Escalera-Maurer, Anaïs Le Rhun, Emily K. Crispell, David Weiss, Hannah K. Ratner, Emmanuelle Charpentier, Siddharth Jaggavarapu |
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| Přispěvatelé: | HZI,Helmholtz-Zentrum für Infektionsforschung GmbH, Inhoffenstr. 7,38124 Braunschweig, Germany. |
| Rok vydání: | 2019 |
| Předmět: |
Small RNA
bacterial pathogenesis Transcription Genetic Lipoproteins Virulence Biology medicine.disease_cause Article 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Transcription (biology) CRISPR-Associated Protein 9 medicine CRISPR Francisella novicida DNA Cleavage Francisella Cas9 Molecular Biology Gene 030304 developmental biology Regulation of gene expression Trans-activating crRNA 0303 health sciences DNA Gene Expression Regulation Bacterial Cell Biology Cell biology Protospacer adjacent motif Regulon chemistry RNA CRISPR-Cas Systems gene regulation transcription 030217 neurology & neurosurgery |
| Zdroj: | Molecular cell Mol Cell |
| ISSN: | 1556-5068 |
| DOI: | 10.2139/ssrn.3372971 |
| Popis: | Beyond defense against foreign DNA, the CRISPR-Cas9 system of pathogenic Francisella novicida represses expression of an endogenous immunostimulatory lipoprotein and is essential for virulence. We investigated the specificity and molecular mechanism of this regulation, demonstrating that Cas9 has a highly specific regulon of four genes which must be repressed for bacterial virulence. Regulation occurs through a PAM-dependent interaction of Cas9 with its endogenous DNA targets, directed by a non-canonical small RNA (scaRNA) duplexed with tracrRNA. The limited complementarity between scaRNA and the endogenous DNA targets precludes cleavage. This highlights the evolution of the scaRNA to direct transcriptional interference via interaction with endogenous DNA without lethally targeting the chromosome. We show that scaRNA can be reprogrammed to repress other genes, and with engineered, extended complementarity to an exogenous target, the repurposed scaRNA:tracrRNA-Cas9 machinery can also be licensed to direct cleavage of target DNA. Natural Cas9 transcriptional interference likely represents a broad paradigm of regulatory functionality, which is potentially critical to the physiology of numerous Cas9-encoding pathogenic and commensal organisms. |
| Databáze: | OpenAIRE |
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