Crystal Structure of Staphylococcus aureus Cas9
| Autor: | Yinqing Li, Arisa Kurabayashi, Winston X. Yan, Osamu Nureki, Ryuichiro Ishitani, Feng Zhang, F. Ann Ran, Hiroshi Nishimasu, Bernd Zetsche, Le Cong |
|---|---|
| Přispěvatelé: | Institute for Medical Engineering and Science, Massachusetts Institute of Technology. Department of Biological Engineering, Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences, McGovern Institute for Brain Research at MIT, Cong, Le, Yan, Winston Xia, Zetsche, Bernd, Li, Yinqing, Zhang, Feng |
| Rok vydání: | 2015 |
| Předmět: |
Models
Molecular Staphylococcus aureus Streptococcus pyogenes Molecular Sequence Data Sequence alignment Biology Crystallography X-Ray Article General Biochemistry Genetics and Molecular Biology chemistry.chemical_compound Bacterial Proteins Genome editing Amino Acid Sequence Guide RNA Subgenomic mRNA Genetics Biochemistry Genetics and Molecular Biology(all) Cas9 RNA DNA Protein Structure Tertiary 3. Good health Protospacer adjacent motif chemistry CRISPR-Cas Systems Genetic Engineering Sequence Alignment RNA Guide Kinetoplastida |
| Zdroj: | PMC |
| ISSN: | 0092-8674 |
| DOI: | 10.1016/j.cell.2015.08.007 |
| Popis: | Summary The RNA-guided DNA endonuclease Cas9 cleaves double-stranded DNA targets with a protospacer adjacent mot if (PAM) and complementarity to the guide RNA. Recently, we harnessed Staphylococcus aureus Cas9 (SaCas9), which is significantly smaller than Streptococcus pyogenes Cas9 (SpCas9), to facilitate efficient in vivo genome editing. Here, we report the crystal structures of SaCas9 in complex with a single guide RNA (sgRNA) and its double-stranded DNA targets, containing the 5′-TTGAAT-3′ PAM and the 5′-TTGGGT-3′ PAM, at 2.6 and 2.7 Å resolutions, respectively. The structures revealed the mechanism of the relaxed recognition of the 5′-NNGRRT-3′ PAM by SaCas9. A structural comparison of SaCas9 with SpCas9 highlighted both structural conservation and divergence, explaining their distinct PAM specificities and orthologous sgRNA recognition. Finally, we applied the structural information about this minimal Cas9 to rationally design compact transcriptional activators and inducible nucleases, to further expand the CRISPR-Cas9 genome editing toolbox. National Institute of General Medical Sciences (U.S.) (Grant T32GM007753) National Institutes of Health (U.S.) (Award 1DP1-MH100706) |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|