Methods and compositions for assessing CRISPER/Cas-mediated disruption or excision and CRISPR/Cas-induced recombination with an exogenous donor nucleic acid in vivo

US11021719B2 · kind B2 · utility

Assignee

• REGENERON PHARMACEUTICALS, INC. · US

Inventors

• Guochun Gong · Pleasantville, US
• Charleen Hunt · Dumont, US
• Susannah Brydges · Scarsdale, US
• Suzanne Hartford · Putnam Valley, US
• David Frendewey · New York, US
• Brian Zambrowicz · Diemelstraße, US
• Andrew J. Murphy · Croton-on-Hudson, US

Key dates

• Filing date: Jul 31, 2018
• Grant date: Jun 1, 2021
• Expiry date: Dec 12, 2038

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12Y302/01023
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

Methods and compositions are provided for assessing CRISPR/Cas-mediated non-homologous end joining (NHEJ) activity and/or CRISPR/Cas-induced recombination of a target genomic locus with an exogenous donor nucleic acid in vivo or ex vivo. The methods and compositions employ non-human animals comprising a CRISPR reporter such as a genomically integrated CRISPR reporter for detecting and measuring targeted excision of a sequence between two CRISPR/Cas nuclease cleavage sites or disruption of a sequence near a CRISPR/Cas nuclease cleavage site and/or measuring CRISPR/Cas-induced recombination of the CRISPR reporter with an exogenous donor nucleic acid to convert the coding sequence for a first reporter protein to the coding sequence for a different second reporter protein. Methods and compositions are also provided for making and using these non-human animals.