Precise Cas9 targeting enables genomic mutation prevention
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Author(s)
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
Date Issued
March 2018
Journal
Proceedings of the National Academy of Sciences
Publisher
National Academy of Sciences (U.S.)
Citation
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.
Version
Final published version
Abstract
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.
MIT Department
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)
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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.
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DOI of Published Version
https://doi.org/10.1073/pnas.1718148115
