puc-header

Catalytically Active Cas9 Mediates Transcriptional Interference to Facilitate Bacterial Virulence

62 Pages Posted: 16 Apr 2019 Sneak Peek Status: Review Complete

See all articles by Hannah K. Ratner

Hannah K. Ratner

Emory University - Microbiology and Molecular Genetics Program; Emory University - Emory Vaccine Center; Emory University - Yerkes National Primate Research Center

Andrés Escalera-Maurer

Max Planck Unit for the Science of Pathogens; Helmholtz Centre for Infection Research

Anaïs Le Rhun

Max Planck Unit for the Science of Pathogens; Helmholtz Centre for Infection Research

Siddarth Jaggavarapu

Emory University - Emory Vaccine Center; Emory University - Yerkes National Primate Research Center; Emory University - Division of Infectious Diseases

Jessie E. Wozniak

Emory University - Microbiology and Molecular Genetics Program; Emory University - Emory Vaccine Center; Emory University - Yerkes National Primate Research Center

Emily K. Crispell

Emory University - Microbiology and Molecular Genetics Program; Emory University - Emory Vaccine Center; Emory University - Yerkes National Primate Research Center

Emmanuelle Charpentier

Max Planck Unit for the Science of Pathogens; Helmholtz Centre for Infection Research

David S. Weiss

Emory University - Microbiology and Molecular Genetics Program; Emory University - Emory Vaccine Center; Emory University - Yerkes National Primate Research Center; Emory University - Division of Infectious Diseases

More...

Abstract

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.

Suggested Citation

Ratner, Hannah K. and Escalera-Maurer, Andrés and Le Rhun, Anaïs and Jaggavarapu, Siddarth and Wozniak, Jessie E. and Crispell, Emily K. and Charpentier, Emmanuelle and Weiss, David S., Catalytically Active Cas9 Mediates Transcriptional Interference to Facilitate Bacterial Virulence (April 16, 2019). Available at SSRN: https://ssrn.com/abstract=3372971 or http://dx.doi.org/10.2139/ssrn.3372971
This is a paper under consideration at Cell Press and has not been peer-reviewed.

Hannah K. Ratner

Emory University - Microbiology and Molecular Genetics Program

United States

Emory University - Emory Vaccine Center

United States

Emory University - Yerkes National Primate Research Center

United States

Andrés Escalera-Maurer

Max Planck Unit for the Science of Pathogens

Germany

Helmholtz Centre for Infection Research

Inhoffenstraße 7
Braunschweig, 38124
Germany

Anaïs Le Rhun

Max Planck Unit for the Science of Pathogens

Germany

Helmholtz Centre for Infection Research

Inhoffenstraße 7
Braunschweig, 38124
Germany

Siddarth Jaggavarapu

Emory University - Emory Vaccine Center

United States

Emory University - Yerkes National Primate Research Center

United States

Emory University - Division of Infectious Diseases

United States

Jessie E. Wozniak

Emory University - Microbiology and Molecular Genetics Program

United States

Emory University - Emory Vaccine Center

United States

Emory University - Yerkes National Primate Research Center

United States

Emily K. Crispell

Emory University - Microbiology and Molecular Genetics Program

United States

Emory University - Emory Vaccine Center

United States

Emory University - Yerkes National Primate Research Center

United States

Emmanuelle Charpentier

Max Planck Unit for the Science of Pathogens

Germany

Helmholtz Centre for Infection Research

Inhoffenstraße 7
Braunschweig, 38124
Germany

David S. Weiss (Contact Author)

Emory University - Microbiology and Molecular Genetics Program ( email )

United States

Emory University - Emory Vaccine Center ( email )

United States

Emory University - Yerkes National Primate Research Center ( email )

United States

Emory University - Division of Infectious Diseases ( email )

United States

Click here to go to Cell.com

Go to Cell.com

Paper statistics

Abstract Views
195
PlumX Metrics
Downloads
10