Precise Cas9 targeting enables genomic mutation prevention
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; ... Show more Show less
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Show full item recordAbstract
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.
Date issued
2018-03Department
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)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
ISSN
0027-8424
1091-6490