Crystal Structure of the Minimal Cas9 from Campylobacter jejuni Reveals the Molecular Diversity in the CRISPR-Cas9 Systems
The RNA-guided endonuclease Cas9 generates a double-strand break at DNA target sites complementary to the guide RNA and has been harnessed for the development of a variety of new technologies, such as genome editing. Here, we report the crystal structures of Campylobacter jejuni Cas9 (CjCas9), one o...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | en_US |
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Elsevier
2018
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| Online Access: | http://hdl.handle.net/1721.1/114953 https://orcid.org/0000-0003-2782-2509 |
| _version_ | 1811096514034925568 |
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| author | Yamada, Mari Watanabe, Yuto Hirano, Hisato Ran, F. Ann Nakane, Takanori Ishitani, Ryuichiro Nishimasu, Hiroshi Nureki, Osamu Gootenberg, Jonathan S Zhang, Feng |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Yamada, Mari Watanabe, Yuto Hirano, Hisato Ran, F. Ann Nakane, Takanori Ishitani, Ryuichiro Nishimasu, Hiroshi Nureki, Osamu Gootenberg, Jonathan S Zhang, Feng |
| author_sort | Yamada, Mari |
| collection | MIT |
| description | The RNA-guided endonuclease Cas9 generates a double-strand break at DNA target sites complementary to the guide RNA and has been harnessed for the development of a variety of new technologies, such as genome editing. Here, we report the crystal structures of Campylobacter jejuni Cas9 (CjCas9), one of the smallest Cas9 orthologs, in complex with an sgRNA and its target DNA. The structures provided insights into a minimal Cas9 scaffold and revealed the remarkable mechanistic diversity of the CRISPR-Cas9 systems. The CjCas9 guide RNA contains a triple-helix structure, which is distinct from known RNA triple helices, thereby expanding the natural repertoire of RNA triple helices. Furthermore, unlike the other Cas9 orthologs, CjCas9 contacts the nucleotide sequences in both the target and non-target DNA strands and recognizes the 5′-NNNVRYM-3′ as the protospacer-adjacent motif. Collectively, these findings improve our mechanistic understanding of the CRISPR-Cas9 systems and may facilitate Cas9 engineering. Keywords: CRISPR-Cas system; Cas9; protospacer adjacent motif; RNA triplex; crystal structure |
| first_indexed | 2024-09-23T16:44:53Z |
| format | Article |
| id | mit-1721.1/114953 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T16:44:53Z |
| publishDate | 2018 |
| publisher | Elsevier |
| record_format | dspace |
| spelling | mit-1721.1/1149532022-09-29T21:14:54Z Crystal Structure of the Minimal Cas9 from Campylobacter jejuni Reveals the Molecular Diversity in the CRISPR-Cas9 Systems Yamada, Mari Watanabe, Yuto Hirano, Hisato Ran, F. Ann Nakane, Takanori Ishitani, Ryuichiro Nishimasu, Hiroshi Nureki, Osamu Gootenberg, Jonathan S Zhang, Feng 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 Zhang, Feng Gootenberg, Jonathan S Zhang, Feng The RNA-guided endonuclease Cas9 generates a double-strand break at DNA target sites complementary to the guide RNA and has been harnessed for the development of a variety of new technologies, such as genome editing. Here, we report the crystal structures of Campylobacter jejuni Cas9 (CjCas9), one of the smallest Cas9 orthologs, in complex with an sgRNA and its target DNA. The structures provided insights into a minimal Cas9 scaffold and revealed the remarkable mechanistic diversity of the CRISPR-Cas9 systems. The CjCas9 guide RNA contains a triple-helix structure, which is distinct from known RNA triple helices, thereby expanding the natural repertoire of RNA triple helices. Furthermore, unlike the other Cas9 orthologs, CjCas9 contacts the nucleotide sequences in both the target and non-target DNA strands and recognizes the 5′-NNNVRYM-3′ as the protospacer-adjacent motif. Collectively, these findings improve our mechanistic understanding of the CRISPR-Cas9 systems and may facilitate Cas9 engineering. Keywords: CRISPR-Cas system; Cas9; protospacer adjacent motif; RNA triplex; crystal structure United States. Department of Energy (Grant DE-FG02-97ER25308) National Institute of Mental Health (U.S.) (Grant 5DP1-MH100706) National Institute of Mental Health (U.S.) (Grant 1R01-MH110049) 2018-04-25T17:38:12Z 2018-04-25T17:38:12Z 2017-03 2017-01 Article http://purl.org/eprint/type/JournalArticle 1097-2765 1097-4164 http://hdl.handle.net/1721.1/114953 Yamada, Mari et al. “Crystal Structure of the Minimal Cas9 from Campylobacter Jejuni Reveals the Molecular Diversity in the CRISPR-Cas9 Systems.” Molecular Cell 65, 6 (March 2017): 1109–1121 © 2017 Elsevier Inc https://orcid.org/0000-0003-2782-2509 en_US https://doi.org/10.1016/j.molcel.2017.02.007 Molecular Cell Creative Commons Attribution-NonCommercial-NoDerivs License http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf Elsevier Prof. Zhang via Courtney Crummett |
| spellingShingle | Yamada, Mari Watanabe, Yuto Hirano, Hisato Ran, F. Ann Nakane, Takanori Ishitani, Ryuichiro Nishimasu, Hiroshi Nureki, Osamu Gootenberg, Jonathan S Zhang, Feng Crystal Structure of the Minimal Cas9 from Campylobacter jejuni Reveals the Molecular Diversity in the CRISPR-Cas9 Systems |
| title | Crystal Structure of the Minimal Cas9 from Campylobacter jejuni Reveals the Molecular Diversity in the CRISPR-Cas9 Systems |
| title_full | Crystal Structure of the Minimal Cas9 from Campylobacter jejuni Reveals the Molecular Diversity in the CRISPR-Cas9 Systems |
| title_fullStr | Crystal Structure of the Minimal Cas9 from Campylobacter jejuni Reveals the Molecular Diversity in the CRISPR-Cas9 Systems |
| title_full_unstemmed | Crystal Structure of the Minimal Cas9 from Campylobacter jejuni Reveals the Molecular Diversity in the CRISPR-Cas9 Systems |
| title_short | Crystal Structure of the Minimal Cas9 from Campylobacter jejuni Reveals the Molecular Diversity in the CRISPR-Cas9 Systems |
| title_sort | crystal structure of the minimal cas9 from campylobacter jejuni reveals the molecular diversity in the crispr cas9 systems |
| url | http://hdl.handle.net/1721.1/114953 https://orcid.org/0000-0003-2782-2509 |
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