Precise Cas9 targeting enables genomic mutation prevention
Here, we present a generalized method of guide RNA “tuning” that enables Cas9 to discriminate between two target sites that differ by a single-nucleotide polymorphism. We employ our methodology to generate an in vivo mutation prevention system in which Cas9 actively restricts the occurrence of undes...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
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National Academy of Sciences (U.S.)
2018
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| Online Access: | http://hdl.handle.net/1721.1/117584 https://orcid.org/0000-0001-8167-7096 https://orcid.org/0000-0002-5560-8246 |
| _version_ | 1826212856427708416 |
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| author | Chavez, Alejandro Pruitt, Benjamin W. Tuttle, Marcelle Cecchi, Ryan J. Winston, Jordan Turczyk, Brian M. Tung, Michael Collins, James J. Church, George M Shapiro, Rebecca |
| author2 | Massachusetts Institute of Technology. Institute for Medical Engineering & Science |
| author_facet | Massachusetts Institute of Technology. Institute for Medical Engineering & Science Chavez, Alejandro Pruitt, Benjamin W. Tuttle, Marcelle Cecchi, Ryan J. Winston, Jordan Turczyk, Brian M. Tung, Michael Collins, James J. Church, George M Shapiro, Rebecca |
| author_sort | Chavez, Alejandro |
| collection | MIT |
| description | Here, we present a generalized method of guide RNA “tuning” that enables Cas9 to discriminate between two target sites that differ by a single-nucleotide polymorphism. We employ our methodology to generate an in vivo mutation prevention system in which Cas9 actively restricts the occurrence of undesired gain-of-function mutations within a population of engineered organisms. We further demonstrate that the system is scalable to a multitude of targets and that the general tuning and prevention concepts are portable across engineered Cas9 variants and Cas9 orthologs. Finally, we show that the mutation prevention system maintains robust activity even when placed within the complex environment of the mouse gastrointestinal tract. |
| first_indexed | 2024-09-23T15:38:53Z |
| format | Article |
| id | mit-1721.1/117584 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T15:38:53Z |
| publishDate | 2018 |
| publisher | National Academy of Sciences (U.S.) |
| record_format | dspace |
| spelling | mit-1721.1/1175842024-03-20T19:50:30Z Precise Cas9 targeting enables genomic mutation prevention Chavez, Alejandro Pruitt, Benjamin W. Tuttle, Marcelle Cecchi, Ryan J. Winston, Jordan Turczyk, Brian M. Tung, Michael Collins, James J. Church, George M Shapiro, Rebecca Massachusetts Institute of Technology. Institute for Medical Engineering & Science Harvard University--MIT Division of Health Sciences and Technology Massachusetts Institute of Technology. Department of Biological Engineering Program in Media Arts and Sciences (Massachusetts Institute of Technology) Shapiro, Rebecca Sara Collins, James J. Church, George M Here, we present a generalized method of guide RNA “tuning” that enables Cas9 to discriminate between two target sites that differ by a single-nucleotide polymorphism. We employ our methodology to generate an in vivo mutation prevention system in which Cas9 actively restricts the occurrence of undesired gain-of-function mutations within a population of engineered organisms. We further demonstrate that the system is scalable to a multitude of targets and that the general tuning and prevention concepts are portable across engineered Cas9 variants and Cas9 orthologs. Finally, we show that the mutation prevention system maintains robust activity even when placed within the complex environment of the mouse gastrointestinal tract. National Human Genome Research Institute (U.S.) ( Grant P50 HG005550) Wyss Institute for Biologically Inspired Engineering United States. Defense Threat Reduction Agency (Grant HDTRA1-15-1-0051) Paul G. Allen Frontiers Group 2018-08-28T15:43:41Z 2018-08-28T15:43:41Z 2018-03 2018-08-27T17:44:52Z Article http://purl.org/eprint/type/JournalArticle 0027-8424 1091-6490 http://hdl.handle.net/1721.1/117584 Chavez, Alejandro, Benjamin W. Pruitt, Marcelle Tuttle, Rebecca S. Shapiro, Ryan J. Cecchi, Jordan Winston, Brian M. Turczyk, Michael Tung, James J. Collins, and George M. Church. “Precise Cas9 Targeting Enables Genomic Mutation Prevention.” Proceedings of the National Academy of Sciences 115, no. 14 (March 19, 2018): 3669–3673. https://orcid.org/0000-0001-8167-7096 https://orcid.org/0000-0002-5560-8246 http://dx.doi.org/10.1073/pnas.1718148115 Proceedings of the National Academy of Sciences 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 National Academy of Sciences (U.S.) PNAS |
| spellingShingle | Chavez, Alejandro Pruitt, Benjamin W. Tuttle, Marcelle Cecchi, Ryan J. Winston, Jordan Turczyk, Brian M. Tung, Michael Collins, James J. Church, George M Shapiro, Rebecca Precise Cas9 targeting enables genomic mutation prevention |
| title | Precise Cas9 targeting enables genomic mutation prevention |
| title_full | Precise Cas9 targeting enables genomic mutation prevention |
| title_fullStr | Precise Cas9 targeting enables genomic mutation prevention |
| title_full_unstemmed | Precise Cas9 targeting enables genomic mutation prevention |
| title_short | Precise Cas9 targeting enables genomic mutation prevention |
| title_sort | precise cas9 targeting enables genomic mutation prevention |
| url | http://hdl.handle.net/1721.1/117584 https://orcid.org/0000-0001-8167-7096 https://orcid.org/0000-0002-5560-8246 |
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