IP Library › Granted Patent US 12,460,203
Granted Patent B2
US 12,460,203 · App. 17/521,617 · Granted Nov 4, 2025

CRISPR RNA targeting enzymes and systems and uses thereof

Inventors: David R. Cheng (Boston, MA); David A. Scott (San Francisco, CA); Winston X. Yan (Brookline, MA); Shaorong Chong (Arlington, MA)
Assignee: Arbor Biotechnologies, Inc.
C12N15/11C07K14/195C12N9/22C12N15/111C12N15/113C12N15/62C12N2310/20
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Quick Facts
Patent No.
US 12,460,203
App. No.
17/521,617
Granted
Nov 4, 2025
Kind
B2
Abstract

The disclosure describes novel systems, methods, and compositions for the manipulation of nucleic acids in a targeted fashion. The disclosure describes non-naturally occurring, engineered CRISPR systems, components, and methods for targeted modification of a nucleic acid.

Claims (27)

1 . An engineered, non-naturally occurring Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated (Cas) system comprising:

(a) an RNA guide or a nucleic acid encoding the RNA guide comprising a direct repeat sequence of SEQ ID NO: 65 and a spacer sequence capable of hybridizing to a target nucleic acid in a eukaryotic cell; and

(b) a nucleic acid encoding a CRISPR-Cas effector protein, or the CRISPR-Cas effector protein,

wherein the CRISPR-Cas effector protein binds to the RNA guide, and wherein the spacer sequence binds to a target nucleic acid.

2 . The system of claim 1 , wherein the direct repeat consists of an RNA transcript of the nucleotide sequence set forth in SEQ ID NO: 65.

3 . The system of claim 1 , wherein the RNA guide comprises a truncated direct repeat sequence.

4 . The system of claim 1 , wherein the spacer sequence comprises between 15 and 42 nucleotides in length.

5 . The system of claim 1 , wherein the CRISPR-Cas effector protein comprises at least two HEPN domains.

6 . The system of claim 1 , wherein the CRISPR-Cas effector protein comprises an amino acid sequence with at least 95% identity to SEQ ID NO: 26.

7 . The system of claim 6 , wherein the CRISPR-Cas effector protein comprises the amino acid sequence set forth in SEQ ID NO: 26.

8 . The system of claim 1 , wherein the target nucleic acid is an RNA molecule.

9 . The system of claim 1 , wherein the CRISPR-Cas effector protein cleaves the target nucleic acid.

10 . The system of claim 1 , wherein the CRISPR-Cas effector protein further comprises at least one nuclear localization signal (NLS), at least one nuclear export signal (NES), or at least one NLS and at least one NES.

11 . The system of claim 1 , wherein the CRISPR-Cas effector protein further comprises a peptide tag, a fluorescent protein, a base-editing domain, an RNA methyltransferase, an RNA demethylase, a splicing modifier, a localization factor, or a translation modification factor.

12 . The system of claim 1 , wherein the nucleic acid encoding the CRISPR-Cas effector protein is codon-optimized for expression in a cell.

13 . The system of claim 1 , wherein the nucleic acid encoding the RNA guide is operably linked to a promoter.

14 . The system of claim 1 , wherein the nucleic acid encoding the CRISPR-Cas effector protein is operably linked to a promoter.

15 . The system of claim 1 , wherein the nucleic acid encoding the RNA guide is in a vector.

16 . The system of claim 1 , wherein the nucleic acid encoding the CRISPR-Cas effector protein is in a vector.

17 . The system of claim 1 , wherein the nucleic acid encoding the RNA guide and the nucleic acid encoding the CRISPR-Cas effector protein are in a vector.

18 . The system of claim 15 , wherein the vector is a retroviral vector, a lentiviral vector, a phage vector, an adenoviral vector, an adeno-associated vector, or a herpes simplex vector.

19 . The system of claim 1 , wherein the system is present in a delivery system comprising a nanoparticle, a liposome, an exosome, a microvesicle, or a gene-gun.

20 . A cell comprising the system of claim 1 .

21 . A method of binding the system of claim 1 to the target nucleic acid in a cell comprising:

(a) providing the system; and

(b) delivering the system to the cell,

wherein the cell comprises the target nucleic acid, wherein the CRISPR-Cas effector protein binds to the RNA guide, and wherein the spacer sequence binds to the target nucleic acid.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Mar 24, 2022
From: CHENG, DAVID R.; SCOTT, DAVID A.; YAN, WINSTON X.; CHONG, SHAORONG
To: ARBOR BIOTECHNOLOGIES, INC.
Reel/Frame 059390/0902 →
Continuity (10)
Division 15916274 · Mar 8, 2018
Provisional Application 62635443 · Feb 26, 2018
Provisional Application 62628921 · Feb 9, 2018
Provisional Application 62626679 · Feb 5, 2018
Provisional Application 62619691 · Jan 19, 2018
Provisional Application 62587381 · Nov 16, 2017
Provisional Application 62580880 · Nov 2, 2017
Provisional Application 62572367 · Oct 13, 2017
Provisional Application 62527957 · Jun 30, 2017
Related Publication 20220127603A1 · Apr 28, 2022
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