Toward a molecular mechanism-based prediction of CRISPR-Cas9 targeting effects

The CRISPR-Cas9 system,serving as a powerful genome-editing technology,has revolutionized the life sciences.However,it exhi-bits off-target activities that may present severe problems in clin-ical applications.Although a great number of in silico models have been developed to predict CRISPR targeting efficiency and speci-ficity,they are running into a bottleneck with the lack of a mech-anistic understanding of the on-and off-target activities of Cas9.Therefore,we decipher the targeting activities of Cas9 from a molecular mechanism perspective.We expect that a combination of the features derived from the molecular mechanism of Cas9 with state-of-the-art computational modeling will facilitate the development of a better in silico prediction of CRISPR-Cas9 on-/off-target activities.