期刊
MOLECULAR CELL
卷 65, 期 1, 页码 168-175出版社
CELL PRESS
DOI: 10.1016/j.molcel.2016.11.031
关键词
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资金
- Howard Hughes International Student Research Fellowship
- Rita Allen Scholars Program
- Irma T. Hirschl Award
- Sinsheimer Foundation Award
- NIH Director's New Innovator Award [1DP2AI104556-01]
- National Science Foundation
- Paul Allen Foundation
- Howard Hughes Medical Institute
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.
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