US20250304977
2025-10-02
Chemistry; metallurgy
C12N15/746
The patent application details a system for manipulating genetic sequences using CRISPR-Cas technology. This approach leverages vectors that encode components of a CRISPR complex, facilitating precise gene editing in eukaryotic cells. The system is designed to direct CRISPR complex formation and allows for the introduction of specific mutations, enhancing the ability to study genetic factors related to various biological functions and diseases.
At its core, the system utilizes vectors comprising regulatory elements linked to sequences that enable sequence-specific binding of a CRISPR complex. These vectors may include guide sequences and tracr mate sequences that guide the CRISPR enzyme to target DNA sequences. The invention specifies the use of nuclear localization sequences to enhance CRISPR activity within the nucleus of eukaryotic cells, and it includes various configurations for optimizing expression and activity of the CRISPR components.
The described technology addresses the demand for efficient genome editing tools that are easy to implement and scalable across multiple genomic sites. Unlike previous methods requiring customized proteins, this system uses a single Cas enzyme programmed by RNA, simplifying the process. The invention holds potential for applications in synthetic biology, biotechnology, and medicine by enabling systematic perturbation of genetic elements.
The vector system described can be composed of single or multiple vectors, each potentially carrying different components necessary for CRISPR complex assembly and function. These vectors can be plasmids or viral vectors capable of autonomous replication or integration into host genomes. Expression vectors are specifically designed to facilitate gene expression linked to the CRISPR components.
The invention incorporates regulatory elements such as polymerase promoters to control gene expression levels within host cells. The sequence listing is provided in an electronic XML format, ensuring precise documentation of genetic sequences involved. The application also acknowledges federally sponsored research contributions and incorporates extensive references to prior applications and related documents.