Journal
GENES & DEVELOPMENT
Volume 29, Issue 4, Pages 356-361Publisher
COLD SPRING HARBOR LAB PRESS, PUBLICATIONS DEPT
DOI: 10.1101/gad.257550.114
Keywords
CRISPR; adaptation; Cas9; PAM; Streptococcus thermophilus
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Funding
- National Institutes of Health (NIH) [RO1 GM099876]
- National Institute of General Medical Sciences of the NIH [F32GM108263]
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To acquire the ability to recognize and destroy virus and plasmid invaders, prokaryotic CRISPR-Cas systems capture fragments of DNA within the host CRISPR locus. Our results indicate that the process of adaptation by a Type II-A CRISPR-Cas system in Streptococcus thermophilus requires Cas1, Cas2, and Csn2. Surprisingly, we found that Cas9, previously identified as the nuclease responsible for ultimate invader destruction, is also essential for adaptation. Cas9 nuclease activity is dispensable for adaptation. In addition, our studies revealed extensive, unbiased acquisition of the selftargeting host genome sequence by the CRISPR-Cas system that is masked in the presence of active target destruction.
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