CRISPR-associated transposon systems and components

Inventors

Scott, David A. • Cheng, David R. • Yan, Winston X.

Assignees

Arbor Biotechnologies Inc

Abstract

The disclosure describes novel systems, methods, and compositions for the manipulation of nucleic acids in a targeted fashion. The disclosure describes non-naturally occurring, engineered CRISPR systems, components, and methods for targeted modification of DNA, RNA, and protein substrates. Each system includes one or more protein components and one or more nucleic acid components that together target DNA, RNA, or protein substrates.

Core Innovation

The invention provides an engineered, non-naturally occurring Clustered Interspaced Short Palindromic Repeat (CRISPR)-Cas system comprising a Guide and CRISPR-associated proteins. The Guide includes a direct repeat sequence and a spacer sequence capable of hybridizing to a target nucleic acid, where the Guide is CRISPR RNA (crRNA) or DNA. The system is configured to associate guide-directed recognition with CRISPR-associated transposon-related functions through multiple protein domains.

The CRISPR-associated proteins of the system include an amino acid sequence that is at least 90% identical to SEQ ID NO: 19. The system further requires CRISPR-associated proteins comprising an rve integrase domain, a TniQ domain, and a TniB domain. These domains collectively define the engineered CRISPR-associated transposon architecture described for the non-naturally occurring system.

The disclosure describes computational identification of novel CRISPR-Cas loci and definition of minimal system elements as part of establishing engineered CRISPR-associated transposon systems. The system is framed as a CLUST.004377 CRISPR-associated transposon system with CRISPR guide targeting a DNA/RNA substrate and transposase modules enabling DNA cleavage and payload insertion/excision/mobilization, together with additional components associated with integrase and transposon-associated functions and with guidance/target matching and modification options.

Claims Coverage

The provided claims contain one independent claim (clm-00001). It defines an engineered, non-naturally occurring CRISPR-Cas system with a guide capable of hybridizing to a target nucleic acid and a specific set of CRISPR-associated proteins characterized by SEQ ID-based identity and domain composition. The dependent claims further refine delivery, sequence identity constraints, payload structure, and functional outcomes such as insertion/excision/deletion and scarless DNA insertion.

Overall, the claim coverage centers on an engineered, non-naturally occurring CRISPR-Cas system defined by a hybridizing guide (crRNA or DNA) and CRISPR-associated proteins meeting SEQ ID NO: 19 identity plus inclusion of rve integrase, TniQ, and TniB domains; dependent claims add further narrowing via additional identity constraints and payload-related structural elements.

Stated Advantages

Documented Applications