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US 2022/0033791 A1Published application
Enzymes with ruvc domains
AssigneeMetagenomi IP Technologies, LLC
InventorsBrian THOMAS, Christopher BROWN, Rose KANTOR, Audra DEVOTO, Cristina BUTTERFIELD, Lisa ALEXANDER, Daniela S.A. GOLTSMAN, Jason LIU
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Abstract
The present disclosure provides for endonuclease enzymes having distinguishing domain features, as well as methods of using such enzymes or variants thereof.
Claims24
An engineered nuclease system, comprising:
- (a) an endonuclease comprising a RuvC_III domain and an HNH domain;
- and
- (b) an engineered guide ribonucleic acid structure configured to form a complex with said endonuclease comprising:
- (i) a guide ribonucleic acid sequence configured to hybridize to a target deoxyribonucleic acid sequence;
- and
- (ii) a tracr ribonucleic acid sequence configured to bind to said endonuclease, wherein said endonuclease comprises a sequence having at least 90% sequence identity to SEQ ID NO:
- 421.
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An engineered composition comprising:
- (a) an endonuclease comprising a RuvC_III domain having at least 90% sequence identity to SEQ ID NO:
- 2242;
- (b) an engineered guide ribonucleic acid structure configured to form a complex with said endonuclease comprising:
- (i) a guide ribonucleic acid sequence configured to hybridize to a target deoxyribonucleic acid sequence;
- and
- (ii) a ribonucleic acid sequence configured to bind said endonuclease;
- and
- (c) a target DNA sequence comprising a protospacer adjacent motif sequence comprising 5′-RGG-3, wherein R denotes A or G.
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A method of modifying a target deoxyribonucleic acid sequence in a cell, comprising introducing to said cell a composition comprising:
- (a) an endonuclease comprising a RuvC_III domain and an HNH domain;
- and
- (b) an engineered guide ribonucleic acid structure configured to form a complex with said endonuclease comprising:
- (iii) a guide ribonucleic acid sequence configured to hybridize to said target deoxyribonucleic acid sequence;
- and
- (iv) a tracr ribonucleic acid sequence configured to bind to said endonuclease, wherein said endonuclease comprises a sequence having at least 90% sequence identity to SEQ ID NO:
- 421.
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Dependencies are inferred from the claim text.