Invention Title:

CRISPR ENZYMES AND SYSTEMS WITH MODIFIED PAM SPECIFICITY

Publication number:

US20260071198

Publication date:

2026-03-12

Section:

Chemistry; metallurgy

Class:

C12N9/22

Inventors:

Feng Zhang 🇺🇸 Cambridge, MA, United States

David Benjamin Turitz COX 🇺🇸 Cambridge, MA, United States

Ian Slaymaker 🇺🇸 Cambridge, MA, United States

Linyi Gao 🇺🇸 Cambridge, MA, United States

Bernd ZETSCHE 🇺🇸 Cambridge, MA, United States

Assignees:

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

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

Smart overview of the Invention

The patent application details innovative systems, methods, and compositions for targeting nucleic acids using CRISPR technology. It focuses on engineered DNA-targeting systems that include a novel CRISPR effector protein, such as Cpf1 mutants, which have altered PAM (Protospacer Adjacent Motif) specificity. These advancements are significant for the precise manipulation of genetic materials, enabling targeted gene editing and expression control.

Background

Advancements in genome sequencing have highlighted the need for precise genome targeting technologies to manipulate genetic variations effectively. Traditional genome-editing techniques have limitations, necessitating the development of new, scalable, and efficient tools. CRISPR-Cas systems, known for their diversity and adaptability, have emerged as promising candidates for genome engineering, especially with the development of novel effector proteins like those in Class 2 CRISPR-Cas systems.

Technical Details

The invention emphasizes the use of Cpf1, a single RNA-guided endonuclease from Class 2 CRISPR-Cas systems. The focus is on mutant Cpf1 polypeptides that can recognize different or shorter PAM sequences compared to wild-type Cpf1. This expanded PAM recognition allows for more versatile applications in gene targeting and editing. The mutants are capable of recognizing various PAM sequences, such as YCV, NYCV, and others, which broadens the scope of genetic sites that can be targeted.

Applications

This technology has wide-ranging applications in genomics and biotechnology. By expanding the PAM recognition repertoire, these Cpf1 mutants enable more precise and diverse targeting of genetic elements. This capability is crucial for advancing research in synthetic biology, medical applications, and understanding genetic diseases. The invention's ability to target specific genomic sites without adverse effects is a significant contribution to the field.

Conclusion

The invention presents a substantial leap forward in CRISPR technology by modifying PAM specificity through engineered Cpf1 mutants. These advancements provide researchers with powerful tools for genome editing, offering new possibilities for genetic research and therapeutic interventions. The detailed sequence listings and comprehensive understanding of PAM recognition expand the potential for precise and efficient genome manipulation.