Invention Title:

CRISPR ENZYMES AND SYSTEMS

Publication number:

US20250250617

Publication date:

2025-08-07

Section:

Chemistry; metallurgy

Class:

C12Q1/6832

Inventors:

Feng Zhang 🇺🇸 Cambridge, MA, United States

Ian Slaymaker 🇺🇸 Cambridge, MA, United States

Omar O. Abudayyeh 🇺🇸 Cambridge, MA, United States

Jonathan S. Gootenberg 🇺🇸 Cambridge, MA, United States

Bernd ZETSCHE 🇺🇸 Cambridge, MA, United States

Assignees:

President and Fellows of Harvard College 🇺🇸 Cambridge, MA, United States

MASSACHUSETTS INSTITUTE OF TECHNOLOGY 🇺🇸 Cambridge, MA, United States

THE BROAD INSTITUTE, INC. 🇺🇸 Cambridge, MA, United States

Applicants:

Massachusetts Institute of Technology 🇺🇸 Cambridge, MA, United States

PRESIDENT AND FELLOWS OF HARVARD COLLEGE 🇺🇸 Cambridge, MA, United States

The Broad Institute, Inc. 🇺🇸 Cambridge, MA, United States

Drawings (4 of 345)

Drawing 02 for CRISPR ENZYMES AND SYSTEMS

Drawing 03 for CRISPR ENZYMES AND SYSTEMS

Drawing 04 for CRISPR ENZYMES AND SYSTEMS

Drawing 05 for CRISPR ENZYMES AND SYSTEMS

Smart overview of the Invention

The patent application outlines novel systems, methods, and compositions for targeting nucleic acids using engineered DNA-targeting systems. These systems incorporate a unique CRISPR effector protein and at least one targeting nucleic acid component, such as guide RNA. The application details methods for creating and utilizing these systems, as well as the resulting products.

Background and Need

With advancements in genome sequencing and analysis, there is an increasing demand for precise genome-targeting technologies. Current genome-editing techniques like zinc fingers and TALEs have limitations, prompting the need for new strategies that are cost-effective, scalable, and capable of targeting multiple genome positions. The CRISPR-Cas system, particularly its diverse protein composition and architecture, offers potential solutions through novel effector proteins.

Technological Innovations

The invention introduces a method to modify sequences at a target locus by delivering a non-naturally occurring composition comprising a Type V CRISPR-Cas effector protein. This protein forms a complex with nucleic acid components to induce modifications, such as strand breaks. The CRISPR effector proteins, particularly Cpf1, are pivotal in this technology.

Methodology

The invention describes delivering Cpf1 effector proteins with nucleic acid components to modify target sequences. This process involves forming complexes that guide modifications through crRNA or guide RNA. The Cpf1 complexes can introduce staggered cuts with 5' overhangs in DNA sequences, enhancing precision in genetic engineering.

Advanced Features

The invention highlights Cpf1 complexes with engineered nucleic acid components that include guide sequences linked to direct repeats with optimized structures. These may incorporate RNA aptamers capable of binding bacteriophage coat proteins like MS2. Such innovations expand the potential applications of CRISPR technologies in genetic research and biotechnology.