Structure and Dynamics of Cas9 HNH Domain Catalytic State

Date

2017-12-08

Authors

Zuo, Zhicheng
Liu, Jin

ORCID

0000-0002-1067-4063 (Liu, Jin)

Journal Title

Journal ISSN

Volume Title

Publisher

Springer Nature

Abstract

The bacterial CRISPR-Cas9 immune system has been harnessed as a powerful and versatile genome-editing tool and holds immense promise for future therapeutic applications. Despite recent advances in understanding Cas9 structures and its functional mechanism, little is known about the catalytic state of the Cas9 HNH nuclease domain, and identifying how the divalent metal ions affect the HNH domain conformational transition remains elusive. A deeper understanding of Cas9 activation and its cleavage mechanism can enable further optimization of Cas9-based genome-editing specificity and efficiency. Using two distinct molecular dynamics simulation techniques, we have obtained a cross-validated catalytically active state of Cas9 HNH domain primed for cutting the target DNA strand. Moreover, herein we demonstrate the essential roles of the catalytic Mg(2+) for the active state formation and stability. Importantly, we suggest that the derived catalytic conformation of the HNH domain can be exploited for rational engineering of Cas9 variants with enhanced specificity.

Description

Citation

Zuo, Z., & Liu, J. (2017). Structure and Dynamics of Cas9 HNH Domain Catalytic State. Scientific reports, 7(1), 17271. https://doi.org/10.1038/s41598-017-17578-6

Rights

© The Author(s) 2017

License

Attribution 4.0 International (CC BY 4.0)