Jun 14, 2016

Precise Cas9 targeting enables genomic mutation prevention

BioRxiv : the Preprint Server for Biology
Alejandro ChavezGeorge M Church

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 a novel 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 designed mutation prevention system maintains robust activity even when placed within the complex environment of the mouse gastrointestinal tract.

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Mentioned in this Paper

In Vivo
Environment
Gene Mutation
Site
Plasmids
Mouse Gastrointestinal Tract
Single Nucleotide Polymorphism
Orthologous Gene
Gene Mutant
Mutation Abnormality

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