US 12,305,204 B2
CRISPR enzymes and systems
Feng Zhang, Cambridge, MA (US); David Arthur Scott, Cambridge, MA (US); Winston Xia Yan, Cambridge, MA (US); Sourav Choudhury, Cambridge, MA (US); and Matthias Heidenreich, Cambridge, MA (US)
Assigned to THE BROAD INSTITUTE, INC., Cambridge, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
Filed by THE BROAD INSTITUTE, INC., Cambridge, MA (US); and MASSACHUSETTS INSTITUTE OF TECHNOLOGY, Cambridge, MA (US)
Filed on Oct. 17, 2023, as Appl. No. 18/488,261.
Application 18/488,261 is a continuation of application No. 16/325,892, previously published as PCT/US2017/047458, filed on Aug. 17, 2017.
Claims priority of provisional application 62/437,031, filed on Dec. 20, 2016.
Claims priority of provisional application 62/376,372, filed on Aug. 17, 2016.
Prior Publication US 2024/0124860 A1, Apr. 18, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 9/22 (2006.01); C12N 15/10 (2006.01); C12N 15/86 (2006.01); G16B 20/00 (2019.01); G16B 30/10 (2019.01); G16B 30/20 (2019.01)
CPC C12N 9/22 (2013.01) [C12N 15/102 (2013.01); C12N 15/1089 (2013.01); C12N 15/86 (2013.01); G16B 20/00 (2019.02); G16B 30/10 (2019.02); G16B 30/20 (2019.02); C12N 2310/20 (2017.05); C12N 2710/10343 (2013.01); C12N 2740/16043 (2013.01); C12N 2750/14143 (2013.01); C12N 2800/80 (2013.01)] 14 Claims
 
1. A method for preparing a Type II or a Type V CRISPR-Cas guide molecule comprising:
selecting a set of candidate therapeutic target sequences for one or more loci in a target population, wherein the candidate therapeutic target sequences do not contain variants occurring above a threshold allele frequency in the target population;
removing any candidate therapeutic target sequences having off-target candidates in haplotypes that occur in at least 0.1% of the target population from the set of candidate therapeutic target sequences to thereby define a final target sequence set; and
preparing one or more guide molecules based on the final therapeutic target sequence set targeting one or more loci associated with a disease or disorder.