Unified energetics analysis unravels SpCas9 cleavage activity for optimal gRNA design.

ABSTRACT: While CRISPR/Cas9 is a powerful tool in genome engineering, the on-target activity and off-target effects of the system widely vary because of the differences in guide RNA (gRNA) sequences and genomic environments. Traditional approaches rely on separate models and parameters to treat on- and off-target cleavage activities. Here, we demonstrate that a free-energy scheme dominates the Cas9 editing efficacy and delineate a method that simultaneously considers on-target activities and off-target effects. While data-driven machine-learning approaches learn rules to model particular datasets, they may not be as transferrable to new systems or capable of producing new mechanistic insights as principled physical approaches. By integrating the energetics of R-loop formation under Cas9 binding, the

SUBMITTER: Zhang D 

PROVIDER: S-EPMC6500161 | biostudies-literature | 2019 Apr

REPOSITORIES: biostudies-literature