Crystal Structure of Staphylococcus aureus Cas9
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 fac...
| Main Authors: | , , , , , , , , , |
|---|---|
| Other Authors: | |
| Format: | Article |
| Published: |
Elsevier
2017
|
| Online Access: | http://hdl.handle.net/1721.1/112724 https://orcid.org/0000-0002-3067-479X https://orcid.org/0000-0003-2782-2509 |
| _version_ | 1826204073873899520 |
|---|---|
| author | Nishimasu, Hiroshi Ran, F. Ann Kurabayashi, Arisa Ishitani, Ryuichiro Nureki, Osamu Cong, Le Yan, Winston Xia Zetsche, Bernd Li, Yinqing Zhang, Feng |
| author2 | Institute for Medical Engineering and Science |
| author_facet | Institute for Medical Engineering and Science Nishimasu, Hiroshi Ran, F. Ann Kurabayashi, Arisa Ishitani, Ryuichiro Nureki, Osamu Cong, Le Yan, Winston Xia Zetsche, Bernd Li, Yinqing Zhang, Feng |
| author_sort | Nishimasu, Hiroshi |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T12:48:30Z |
| format | Article |
| id | mit-1721.1/112724 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T12:48:30Z |
| publishDate | 2017 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1127242022-09-28T10:09:40Z Crystal Structure of Staphylococcus aureus Cas9 Nishimasu, Hiroshi Ran, F. Ann Kurabayashi, Arisa Ishitani, Ryuichiro Nureki, Osamu Cong, Le Yan, Winston Xia Zetsche, Bernd Li, Yinqing Zhang, Feng 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 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) 2017-12-13T14:35:53Z 2017-12-13T14:35:53Z 2015-08 2015-08 2017-12-13T13:52:02Z Article http://purl.org/eprint/type/JournalArticle 0092-8674 1097-4172 http://hdl.handle.net/1721.1/112724 Nishimasu, Hiroshi et al. “Crystal Structure of Staphylococcus Aureus Cas9.” Cell 162, 5 (August 2015): 1113–1126 © 2015 Elsevier https://orcid.org/0000-0002-3067-479X https://orcid.org/0000-0003-2782-2509 http://dx.doi.org/10.1016/J.CELL.2015.08.007 Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier PMC |
| spellingShingle | Nishimasu, Hiroshi Ran, F. Ann Kurabayashi, Arisa Ishitani, Ryuichiro Nureki, Osamu Cong, Le Yan, Winston Xia Zetsche, Bernd Li, Yinqing Zhang, Feng Crystal Structure of Staphylococcus aureus Cas9 |
| title | Crystal Structure of Staphylococcus aureus Cas9 |
| title_full | Crystal Structure of Staphylococcus aureus Cas9 |
| title_fullStr | Crystal Structure of Staphylococcus aureus Cas9 |
| title_full_unstemmed | Crystal Structure of Staphylococcus aureus Cas9 |
| title_short | Crystal Structure of Staphylococcus aureus Cas9 |
| title_sort | crystal structure of staphylococcus aureus cas9 |
| url | http://hdl.handle.net/1721.1/112724 https://orcid.org/0000-0002-3067-479X https://orcid.org/0000-0003-2782-2509 |
| work_keys_str_mv | AT nishimasuhiroshi crystalstructureofstaphylococcusaureuscas9 AT ranfann crystalstructureofstaphylococcusaureuscas9 AT kurabayashiarisa crystalstructureofstaphylococcusaureuscas9 AT ishitaniryuichiro crystalstructureofstaphylococcusaureuscas9 AT nurekiosamu crystalstructureofstaphylococcusaureuscas9 AT congle crystalstructureofstaphylococcusaureuscas9 AT yanwinstonxia crystalstructureofstaphylococcusaureuscas9 AT zetschebernd crystalstructureofstaphylococcusaureuscas9 AT liyinqing crystalstructureofstaphylococcusaureuscas9 AT zhangfeng crystalstructureofstaphylococcusaureuscas9 |