IP Library › Granted Patent US 12662667
Granted Patent B2
US 12662667 · App. 17/246,460 · Granted Jun 23, 2026

CRISPR enzymes and systems

Inventors: Eugene Koonin (Bethesda, MD); Feng Zhang (Cambridge, MA); Yuri I. Wolf (Bethesda, MD); Sergey Shmakov (Moscow, RU); Konstantin Severinov (New Brunswick, NJ); Ekaterina Semenova (New Brunswick, NJ); Leonid Minakhin (New Brunswick, MD); Kira S. Makarova (Bethesda, MD); Silvana Konermann (Cambridge, MA); Julia Joung (Cambridge, MA); Jonathan S. Gootenberg (Cambridge, MA); Omar O. Abudayyeh (Cambridge, MA)
Assignees: The Broad Institute, Inc.; Massachusetts Institute of Technology; President and Fellows of Harvard College; Rutgers, the State University of New Jersey; The United States of America, as represented by the Secretary, Department of Health and Human Services; Skolkovo Institute of Science and Technology
C12N15/102A61K48/005C12N9/22C12N15/11C12N15/8213C12N2310/20C12N2800/80
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12662667
App. No.
17/246,460
Granted
Jun 23, 2026
Kind
B2
Abstract

The invention provides for systems, methods, and compositions for targeting nucleic acids. In particular, the invention provides non-naturally occurring or engineered DNA or RNA-targeting systems comprising a novel DNA or RNA-targeting CRISPR effector protein and at least one targeting nucleic acid component like a guide RNA.

Claims (23)

1 . A formed CRISPR-Cas complex, consisting of one or more engineered heterologous nucleic acid components and a Type V Cas polypeptide consisting essentially of a nuclease lobe consisting of an RuvC domain and a heterologous polypeptide functional domain linked to the Type V Cas polypeptide; wherein the RuvC domain is connected to the nuclease lobe by one or more bridge helices, wherein the Type V Cas polypeptide comprises an arginine rich cluster and an inactive zinc finger region, wherein the one or more engineered heterologous nucleic acid components comprise a tracr sequence and direct sequence-specific binding of said CRISPR-Cas complex to a target sequence of a polynucleotide, and wherein the Type V Cas polypeptide does not comprise an HNH domain.

2 . The formed CRISPR-Cas complex of claim 1 , wherein the CRISPR-Cas complex directs cleavage at the target sequence of the polynucleotide.

3 . The formed CRISPR-Cas complex of claim 1 , wherein the one or more engineered heterologous nucleic acid components are engineered to hybridize with a target sequence adjacent to a protospacer motif (PAM) in a genome of a eukaryotic cell.

4 . A vector system comprising one or more viral vectors, wherein the one or more viral vectors encode the Type V Cas polypeptide and the one or more engineered heterologous nucleic acid components of claim 1 .

5 . The vector system of claim 4 , wherein a polynucleotide sequence in the one or more viral vectors encoding the Type V Cas polypeptide is codon optimized for expression in a eukaryotic cell.

6 . The vector system of claim 4 , wherein the one or more engineered heterologous nucleic acid component comprises two separate components, a direct repeat component and a spacer component.

7 . The vector system of claim 4 , wherein the one or more engineered heterologous nucleic acid components are engineered to hybridize with a target sequence adjacent to a protospacer motif (PAM) in a genome of a eukaryotic cell.

8 . The vector system of claim 4 , wherein the viral vector is an adenoviral vector, a lentiviral vector, or an adeno-associated viral vector.

9 . The vector system of claim 4 , wherein the one or more engineered heterologous nucleic acid component comprises two separate components, a direct repeat component and a spacer component.

10 . The formed CRISPR-Cas complex of claim 1 , comprising an mRNA encoding the Cas polypeptide, wherein the mRNA is comprised in a lipid nanoparticle, a liposome, an exosome, or a microvesicle.

11 . The formed CRISPR-Cas complex of claim 1 , wherein the functional domain is a deaminase.

12 . The formed CRISPR-Cas complex of claim 1 , wherein the one or more engineered heterologous nucleic acid component comprises two separate components, a direct repeat component and a spacer component.

13 . The formed CRISPR-Cas complex of claim 1 , wherein the heterologous nucleic acid component comprises two separate components, a direct repeat component and a spacer component.

14 . The formed CRISPR-Cas complex of claim 1 , comprising one or more viral vectors encoding the Type V Cas polypeptide, wherein the viral vector is an adenoviral vector, a lentiviral vector, or an adeno-associated viral vector.

15 . A method of targeting a polynucleotide, comprising contacting a sample comprising a polynucleotide with a CRISPR-Cas complex consisting of 1) a Type V Cas polypeptide consisting essentially of a nuclease lobe consisting of an RuvC domain and a heterologous polypeptide functional domain linked to the Type V Cas polypeptide, wherein the RuvC domain is connected to the nuclease lobe by one or more bridge helices, wherein the Type V Cas polypeptide comprises an arginine rich cluster, and an inactive zinc finger, wherein the Type V Cas polypeptide does not comprise an HNH domain, and 2) one or more engineered heterologous nucleic acid components comprise a tracr sequence and are capable of directing sequence-specific binding of the CRISPR-Cas complex to a target sequence of the polynucleotide.

16 . The method of claim 15 , wherein the target sequence is in a eukaryotic cell.

17 . The method of claim 16 , wherein contacting the eukaryotic cell with the CRISPR-Cas complex results in modification of a gene or gene product, or modification in an expression of a gene product.

18 . The method of claim 15 , wherein contacting an eukaryotic cell with the CRISPR-Cas complex results in cleavage of the target sequence.

19 . The method of claim 15 , wherein the one or more engineered heterologous nucleic acid components comprise two separate components, a direct repeat component and a spacer component.

20 . A method of targeting a polynucleotide, comprising contacting a sample comprising, 1) a catalytically inactive Type V Cas polypeptide linked to one or more heterologous polypeptide functional domains, wherein the Type V Cas polypeptide consists essentially a nuclease lobe consisting of a mutated RuvC domain, wherein the Type V Cas polypeptide does not comprise an HNH domain, wherein the mutated RuvC domain is connected to the nuclease lobe by one or more bridge helices, wherein the Type V Cas polypeptide comprises an arginine rich cluster, and an inactive zinc finger, and 2) one or more heterologous engineered nucleic acid components that comprise a tracr sequence and that form a CRISPR-Cas complex with the Cas polypeptide and that is capable of directing sequence-specific binding of said complex to a target sequence of a polynucleotide.

21 . The method of claim 20 , wherein the target sequence is in a eukaryotic cell.

22 . The method of claim 21 , wherein contacting the eukaryotic cell with the CRISPR-Cas complex results in modification of a gene or gene product, or modification in expression of a gene product.

23 . The method of claim 20 , wherein the one or more engineered heterologous nucleic acid components comprise two separate components, a direct repeat component and a spacer component.