The patent badge is an abbreviated version of the USPTO patent document. The patent badge does contain a link to the full patent document.
The patent badge is an abbreviated version of the USPTO patent document. The patent badge covers the following: Patent number, Date patent was issued, Date patent was filed, Title of the patent, Applicant, Inventor, Assignee, Attorney firm, Primary examiner, Assistant examiner, CPCs, and Abstract. The patent badge does contain a link to the full patent document (in Adobe Acrobat format, aka pdf). To download or print any patent click here.
Patent No.:
Date of Patent:
Dec. 13, 2022
Filed:
Feb. 14, 2018
Universita' Degli Studi Di Trento, Trento, IT;
Anna Cereseto, Trento, IT;
Antonio Casini, Pagnacco, IT;
Gianluca Petris, Sauris, IT;
Alberto Inga, Trento, IT;
Michele Olivieri, Toronto, CA;
UNIVERSITA' DEGLI STUDI DI TRENTO, Trento, IT;
Abstract
To address the limitations deriving from the unspecific genomic cleavages of theCas9 (SpCas9) and to identify variants with higher cleavage fidelity, the present invention describes a yeast-based assay which allows to simultaneously evaluate the on- and off-target activity towards two engineered genomic targets. The screening of SpCas9 variants obtained by random mutagenesis of the Red-II domain allowed the identification of hits with increased on/off ratios. The best performing nuclease, evoCas9, was isolated through the combination of the identified mutations within a single variant. Side by side analyses with previously reported rationally designed variants demonstrated a significant improvement in fidelity of evoCas9 of the present invention.