	US 12,467,045 B2
	Tuning CRISPR/Cas9 activity with chemically modified nucleotide substitutions
Keith Gagnon, Carbondale, IL (US); Masad Damha, Montreal (CA); Elise Malek-Adamian, Ottawa (CA); Maryam Habibian, Montreal (CA); Daniel O'Reilly, Waterlooville (GB); and Zachary Kartje, Valparaiso, IN (US)
Assigned to Board of Trustees of Southern Illinois University, Carbondale, IL (US); and The Royal Institution for the Advancement of Learning/McGill University, Montreal (CA)
Filed by The Royal Institution for the Advancement of Learning/McGill University, Montreal (CA); and Board of Trustees of Southern Illinois University, Carbondale, IL (US)
Filed on Mar. 18, 2022, as Appl. No. 17/698,390.
Application 17/698,390 is a division of application No. 16/327,605, granted, now 11,306,305, previously published as PCT/US2017/053041, filed on Sep. 22, 2017.
Claims priority of provisional application 62/398,775, filed on Sep. 23, 2016.
Prior Publication US 2024/0229010 A1, Jul. 11, 2024
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/10 (2006.01); A61K 48/00 (2006.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12N 15/113 (2010.01); C12N 15/90 (2006.01)

CPC C12N 15/102 (2013.01) [A61K 48/00 (2013.01); C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12N 15/113 (2013.01); C12N 15/90 (2013.01); C12N 15/907 (2013.01); C12N 2310/20 (2017.05); C12N 2310/315 (2013.01); C12N 2310/318 (2013.01); C12N 2310/319 (2013.01); C12N 2310/3231 (2013.01); C12N 2310/344 (2013.01); C12N 2310/346 (2013.01); C12N 2800/80 (2013.01)]

24 Claims

1. A composition comprising a Cas9 protein, a first oligonucleotide comprising a first region having a 5′ end and a 3′ end and a second region, and a second oligonucleotide comprising a first region and a second region, wherein said second region of said first oligonucleotide interacts with said first region of said second oligonucleotide, and said second region of said second oligonucleotide interacts with said Cas9 protein,

wherein said first oligonucleotide:

a) comprises at least a first 2′-deoxyribonucleotide or 2′-deoxyribonucleotide analog in said second region or within about 12 nucleotides of said 5′ end of said first region; and

b) comprises at least two ribonucleotides or ribonucleotide analogs within about 8 nucleotides of the 3′ end of said first region,

wherein said second region of said first oligonucleotide comprises the chemical structure of crRNA-E2_J.