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US 12,215,315 B2Granted
Method for modifying target nucleic acid in genome of cell
AssigneeIndustry-Academic Cooperation Foundation, Yonsei University
InventorsHyong Bum Kim, Hui Kwon Kim
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Abstract
Provided are: a method for modifying a target nucleic acid in the genome of a cell using a novel PAM sequence; and a cell in which a target nucleic acid in the genome is modified thereby. Accordingly, genome editing can be performed by targeting a position that could not be previously targeted as a target for genome editing, and thus the range of applications of genome editing can be expanded.
Claims4
A method for modifying a target nucleic acid in the genome of a cell, the method comprising:
- incubating a cell comprising a target nucleic acid;
- a polynucleotide encoding a clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas) nuclease or a variant thereof, and a guide RNA,
- wherein the Cas nuclease or a variant thereof is Streptococcus pyogenes Cas9-NG (SpCas9-NG),
- wherein the target nucleic acid comprises a protospacer adjacent motif (PAM) and a target sequence complementary to the guide RNA,
- the PAM consists of a sequence selected from the group consisting of 5′-NTG-3′, 5′-GTH-3′, 5′-VCGD-3′, and 5-TCGDG-3′,
- the target nucleic acid can be recognized by a complex of the Cas nuclease or a variant thereof with the guide RNA, and
- the complex of the Cas nuclease or a variant thereof with the guide RNA modifies the target nucleic acid sequence-specifically.
4 more elements
Dependencies are inferred from the claim text.