Invention Title:

FUNCTIONAL GENOMICS USING CRISPR-CAS SYSTEMS, COMPOSITIONS, METHODS, KNOCK OUT LIBRARIES AND APPLICATIONS THEREOF

Publication number:

US20260218165

Publication date:

2026-07-30

Section:

Chemistry; metallurgy

Class:

C12N15/1034

Inventors:

Feng Zhang 🇺🇸 Cambridge, MA, United States

Neville Espi Sanjana 🇺🇸 Cambridge, MA, United States

Ophir SHALEM 🇺🇸 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 invention pertains to functional genomics utilizing CRISPR-Cas systems, offering compositions and methods for gene function analysis in cells. It encompasses vector systems encoding CRISPR components and techniques for designing and using these vectors. The methods facilitate CRISPR complex formation in eukaryotic cells and leverage the CRISPR-Cas system for functional genomics applications.

Background

Functional genomics aims to understand gene and protein functions and interactions, focusing on dynamic processes like gene transcription and translation rather than static DNA structures. It employs genome-wide approaches and high-throughput methods, contrasting traditional gene-by-gene techniques. The field benefits from genetic screens to elucidate gene roles, pathways, and expression alterations in biological processes.

CRISPR-Cas System

CRISPR-Cas technology enables precise genome editing by programming a Cas enzyme with a short RNA to target specific DNA sequences. This system simplifies genetic factor mapping across diverse biological functions and diseases. Effective use requires understanding engineering, optimization, and tissue-specific delivery of genome editing tools, which are central to this invention.

Genome-Wide Library

The invention includes a genome-wide library of CRISPR-Cas guide RNAs targeting multiple genomic loci to knock out gene functions. This library can target every gene in an organism's genome, applicable to various organisms including eukaryotes, mammals, non-human animals, plants, algae, and fungi. It supports diverse genetic studies and applications.

Gene Knockout Cell Library

A method for generating a gene knockout cell library involves introducing a CRISPR-Cas vector system into cells. This system includes a Cas protein and guide RNAs targeting unique genes, inducing specific genomic cleavage. The resulting library facilitates studying gene functions via knockout mutations. Kits with these libraries and guide RNA panels are available for targeting extensive genomic sequences or specific pathways.