In vivo genome editing using Staphylococcus aureus Cas9
The RNA-guided endonuclease Cas9 has emerged as a versatile genome-editing platform. However, the size of the commonly used Cas9 from Streptococcus pyogenes (SpCas9) limits its utility for basic research and therapeutic applications that use the highly versatile adeno-associated virus (AAV) delivery...
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| Format: | Article |
| Language: | en_US |
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Nature Publishing Group
2016
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| Online Access: | http://hdl.handle.net/1721.1/102586 https://orcid.org/0000-0003-2782-2509 https://orcid.org/0000-0003-1465-1691 https://orcid.org/0000-0002-3067-479X https://orcid.org/0000-0002-2639-9879 https://orcid.org/0000-0003-0369-5269 |
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| author | Ran, F. Ann Cong, Le Yan, Winston X. Gootenberg, Jonathan S. Kriz, Andrea J. Zetsche, Bernd Shalem, Ophir Wu, Xuebing Makarova, Kira S. Koonin, Eugene V. Sharp, Phillip A. Zhang, Feng Yan, Winston Xia Scott, David Arthur |
| author2 | Harvard University--MIT Division of Health Sciences and Technology |
| author_facet | Harvard University--MIT Division of Health Sciences and Technology Ran, F. Ann Cong, Le Yan, Winston X. Gootenberg, Jonathan S. Kriz, Andrea J. Zetsche, Bernd Shalem, Ophir Wu, Xuebing Makarova, Kira S. Koonin, Eugene V. Sharp, Phillip A. Zhang, Feng Yan, Winston Xia Scott, David Arthur |
| author_sort | Ran, F. Ann |
| collection | MIT |
| description | The RNA-guided endonuclease Cas9 has emerged as a versatile genome-editing platform. However, the size of the commonly used Cas9 from Streptococcus pyogenes (SpCas9) limits its utility for basic research and therapeutic applications that use the highly versatile adeno-associated virus (AAV) delivery vehicle. Here, we characterize six smaller Cas9 orthologues and show that Cas9 from Staphylococcus aureus (SaCas9) can edit the genome with efficiencies similar to those of SpCas9, while being more than 1 kilobase shorter. We packaged SaCas9 and its single guide RNA expression cassette into a single AAV vector and targeted the cholesterol regulatory gene Pcsk9 in the mouse liver. Within one week of injection, we observed >40% gene modification, accompanied by significant reductions in serum Pcsk9 and total cholesterol levels. We further assess the genome-wide targeting specificity of SaCas9 and SpCas9 using BLESS, and demonstrate that SaCas9-mediated in vivo genome editing has the potential to be efficient and specific. |
| first_indexed | 2024-09-23T08:08:23Z |
| format | Article |
| id | mit-1721.1/102586 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T08:08:23Z |
| publishDate | 2016 |
| publisher | Nature Publishing Group |
| record_format | dspace |
| spelling | mit-1721.1/1025862022-09-23T11:10:37Z In vivo genome editing using Staphylococcus aureus Cas9 Ran, F. Ann Cong, Le Yan, Winston X. Gootenberg, Jonathan S. Kriz, Andrea J. Zetsche, Bernd Shalem, Ophir Wu, Xuebing Makarova, Kira S. Koonin, Eugene V. Sharp, Phillip A. Zhang, Feng Yan, Winston Xia Scott, David Arthur Harvard University--MIT Division of Health Sciences and Technology Massachusetts Institute of Technology. Department of Biological Engineering Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Brain and Cognitive Sciences McGovern Institute for Brain Research at MIT Koch Institute for Integrative Cancer Research at MIT Cong, Le Kriz, Andrea J. Sharp, Phillip A. Yan, Winston Xia Scott, David Arthur Zhang, Feng Wu, Xuebing The RNA-guided endonuclease Cas9 has emerged as a versatile genome-editing platform. However, the size of the commonly used Cas9 from Streptococcus pyogenes (SpCas9) limits its utility for basic research and therapeutic applications that use the highly versatile adeno-associated virus (AAV) delivery vehicle. Here, we characterize six smaller Cas9 orthologues and show that Cas9 from Staphylococcus aureus (SaCas9) can edit the genome with efficiencies similar to those of SpCas9, while being more than 1 kilobase shorter. We packaged SaCas9 and its single guide RNA expression cassette into a single AAV vector and targeted the cholesterol regulatory gene Pcsk9 in the mouse liver. Within one week of injection, we observed >40% gene modification, accompanied by significant reductions in serum Pcsk9 and total cholesterol levels. We further assess the genome-wide targeting specificity of SaCas9 and SpCas9 using BLESS, and demonstrate that SaCas9-mediated in vivo genome editing has the potential to be efficient and specific. National Institute of General Medical Sciences (U.S.) (T32GM007753) Paul & Daisy Soros Fellowships for New Americans (New York, N.Y.) National Institutes of Health (U.S.) (United States Public Health Service Grant RO1-GM34277) National Institutes of Health (U.S.) (United States Public Health Service Grant R01-CA133404) National Cancer Institute (U.S.) (PO1-CA42063) National Cancer Institute (U.S.) (Cancer Center Support (Core) Grant P30-CA14051) National Institute of Mental Health (U.S.) (5DP1-MH100706) National Institute of Diabetes and Digestive and Kidney Diseases (U.S.) (5R01DK097768-03) National Science Foundation (U.S.) (Waterman Award) W. M. Keck Foundation New York Stem Cell Foundation Damon Runyon Cancer Research Foundation Kinship Foundation. Searle Scholars Program Merkin Foundation Vallee Foundation 2016-05-23T00:19:06Z 2016-05-23T00:19:06Z 2015-04 2014-02 Article http://purl.org/eprint/type/JournalArticle 0028-0836 1476-4687 http://hdl.handle.net/1721.1/102586 Ran, F. Ann, Le Cong, Winston X. Yan, David A. Scott, Jonathan S. Gootenberg, Andrea J. Kriz, Bernd Zetsche, et al. “In Vivo Genome Editing Using Staphylococcus Aureus Cas9.” Nature 520, no. 7546 (April 1, 2015): 186–91. https://orcid.org/0000-0003-2782-2509 https://orcid.org/0000-0003-1465-1691 https://orcid.org/0000-0002-3067-479X https://orcid.org/0000-0002-2639-9879 https://orcid.org/0000-0003-0369-5269 en_US http://dx.doi.org/10.1038/nature14299 Nature Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf Nature Publishing Group PMC |
| spellingShingle | Ran, F. Ann Cong, Le Yan, Winston X. Gootenberg, Jonathan S. Kriz, Andrea J. Zetsche, Bernd Shalem, Ophir Wu, Xuebing Makarova, Kira S. Koonin, Eugene V. Sharp, Phillip A. Zhang, Feng Yan, Winston Xia Scott, David Arthur In vivo genome editing using Staphylococcus aureus Cas9 |
| title | In vivo genome editing using Staphylococcus aureus Cas9 |
| title_full | In vivo genome editing using Staphylococcus aureus Cas9 |
| title_fullStr | In vivo genome editing using Staphylococcus aureus Cas9 |
| title_full_unstemmed | In vivo genome editing using Staphylococcus aureus Cas9 |
| title_short | In vivo genome editing using Staphylococcus aureus Cas9 |
| title_sort | in vivo genome editing using staphylococcus aureus cas9 |
| url | http://hdl.handle.net/1721.1/102586 https://orcid.org/0000-0003-2782-2509 https://orcid.org/0000-0003-1465-1691 https://orcid.org/0000-0002-3067-479X https://orcid.org/0000-0002-2639-9879 https://orcid.org/0000-0003-0369-5269 |
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