IP Library › Granted Patent US 12,286,654
Granted Patent B2
US 12,286,654 · App. 17/841,082 · Granted Apr 29, 2025

Base editing enzymes

Inventors: Jyun-Liang Lin (Emeryville, CA); Alan Brooks (Emeryville, CA); Cristina Butterfield (Emeryville, CA); Christopher Brown (Emeryville, CA); Cindy Castelle (Emeryville, CA); Brian C. Thomas (Emeryville, CA)
Assignee: Metagenomi, Inc.
C12N9/22C12N9/78C12N15/11C12N15/111C12N15/1137C12N15/85C12N15/907C12Y302/02027C12Y305/04004C12N2310/20C12N2800/107C12N2800/80
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Quick Facts
Patent No.
US 12,286,654
App. No.
17/841,082
Granted
Apr 29, 2025
Kind
B2
Abstract

The present disclosure provides for endonuclease enzymes having distinguishing domain features, as well as methods of using such enzymes or variants thereof.

Claims (20)

1. An engineered nucleic acid editing system comprising:

(a) an endonuclease comprising a RuvC domain and an HNH domain, wherein the endonuclease is a class 2, type II Cas endonuclease, wherein the endonuclease comprises the amino acid sequence of any one of SEQ ID NO: 597 or 70-78;

(b) a base editor coupled to the endonuclease, wherein the base editor comprises an amino acid sequence having at least 95% sequence identity to the amino acid sequence of SEQ ID NO: 386, wherein the base editor is an adenosine deaminase; and

(c) an engineered guide ribonucleic acid structure configured to form a complex with the endonuclease, wherein the engineered guide ribonucleic acid structure comprises:

(i) a guide portion configured such that the guide portion hybridizes to a target deoxyribonucleic acid; and

(ii) a non-guide portion configured such that the non-guide portion binds the endonuclease.

2. The engineered nucleic acid editing system of claim 1 , wherein the endonuclease is configured to bind to the target deoxyribonucleic acid, wherein the target deoxyribonucleic acid comprises a protospacer adjacent motif (PAM) sequence comprising SEQ ID NO: 598.

3. The engineered nucleic acid editing system of claim 1 , wherein the engineered guide ribonucleic acid structure comprises a sequence having at least 80% sequence identity to any one of SEQ ID NO: 88-96 or 488-489.

4. The engineered nucleic acid editing system of claim 3 , wherein the engineered guide ribonucleic acid structure comprises a sequence having at least 80% sequence identity to SEQ ID NO: 489.

5. The engineered nucleic acid editing system of claim 1 , wherein the guide portion and the non-guide portion are in a single ribonucleic acid.

6. The engineered nucleic acid editing system of claim 1 , wherein a region of the guide portion is complementary to a eukaryotic, fungal, plant, mammalian, or human genomic target.

7. The engineered nucleic acid editing system of claim 1 , wherein the guide portion is 15-24 nucleotides in length.

8. The engineered nucleic acid editing system of claim 1 , wherein the endonuclease or the base editor further comprises one or more nuclear localization sequences (NLSs).

9. The engineered nucleic acid editing system of claim 1 , wherein the endonuclease is covalently coupled directly to the base editor or covalently coupled to the base editor through a linker.

10. The engineered nucleic acid editing system of claim 9 , wherein the endonuclease is covalently coupled directly to the base editor.

11. The engineered nucleic acid editing system of claim 1 , wherein the endonuclease comprises the amino acid sequence of SEQ ID NO: 597.

12. The engineered nucleic acid editing system of claim 1 , wherein the base editor comprises the amino acid sequence SEQ ID NO: 386.

13. The engineered nucleic acid editing system of claim 12 , wherein the endonuclease comprises the amino acid sequence of any one of SEQ ID NO: 70-78.

14. The engineered nucleic acid editing system of claim 11 , wherein the base editor comprises the amino acid sequence of SEQ ID NO: 386.

15. The engineered nucleic acid editing system of claim 9 , wherein the endonuclease covalently coupled to the base editor comprises the amino acid sequence of SEQ ID NO: 476.

Assignments (2)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 8, 2023
From: THOMAS, BRIAN C.
To: METAGENOMI, INC.
Reel/Frame 063892/0196 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 6, 2022
From: LIN, JYUN-LIANG; BROOKS, ALAN; BUTTERFIELD, CRISTINA; BROWN, CHRISTOPHER; CASTELLE, CINDY
To: METAGENOMI, INC.
Reel/Frame 060997/0835 →
Continuity (4)
Continuation PCTUS2021049962 · Sep 10, 2021
Provisional Application 63222351 · Jul 15, 2021
Provisional Application 63077057 · Sep 11, 2020
Related Publication 20220364067A1 · Nov 17, 2022
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Cited By (1)
US 12,503,710