Structure-Guided Methods Of Cas9-Mediated Genome Engineering

Patent Status

Brief Description

The ability to program Cas9 for DNA cleavage at sites defined by guide RNAs has led to its adoption as a robust and versatile platform for genome engineering.  Whereas there are a number of ongoing successes with using the CRISPR-Cas9 system for genome engineering, there is a need for understanding the structural basis for guide RNA recognition and DNA targeting by Cas9.

UC Berkeley researchers have developed software and methods for providing the structures of Cas9 with and without the polynucleotides bound thereto, and have developed the crystals comprising the Cas9 polypeptides.  Using the atomic coordinates, the software can be used to computationally identify a site for amino acid residue substitution, insertion, or deletion to alter a function or chemical property of a Cas9 polypeptide.

Suggested uses

• Genetic engineering or editing of Cas9 polypeptides
• Controlling of site-specific gene regulation

Publication

Structures of Cas9 endonucleases reveal RNA-mediated conformational activation