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© Antoinette Ryter
Serratia marcescens avec présence de flagelles (cils) péritriches. Famille des Enterobacteriaceae, bacille à Gram négatif, non sporulé, anaérobie facultatif, mobile, parfois encapsulé, pouvant synthétiser un pigment rouge ou rose. Présent dans les végétaux , le sol, et l'eau. A l'origine d'infections nosocomiales et résistant à de nombreux antibiotiques. Image colorisée.
Publication : Molecular Cell

Mutations in Cas9 Enhance the Rate of Acquisition of Viral Spacer Sequences during the CRISPR-Cas Immune Response

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in Molecular Cell - 05 Jan 2017

R Heler, AV Wright, M Vucelja, D Bikard, JA Doudna, LA Marraffini

Molecular Cell 65 (1), 168-175

CRISPR loci and their associated (Cas) proteins encode a prokaryotic immune system that protects against viruses and plasmids. Upon infection, a low fraction of cells acquire short DNA sequences from the invader. These sequences (spacers) are integrated in between the repeats of the CRISPR locus and immunize the host against the matching invader. Spacers specify the targets of the CRISPR immune response through transcription into short RNA guides that direct Cas nucleases to the invading DNA molecules. Here we performed random mutagenesis of the RNA-guided Cas9 nuclease to look for variants that provide enhanced immunity against viral infection. We identified a mutation, I473F, that increases the rate of spacer acquisition by more than two orders of magnitude. Our results highlight the role of Cas9 during CRISPR immunization and provide a useful tool to study this rare process and develop it as a biotechnological application.