Compositions and methods for directing proteins to specific loci in the genome
Inventors
Ostertag, Eric • Yeshi, Tseten • Li, Xianghong
Assignees
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Publication Number
US-11473082-B2
Publication Date
2022-10-18
Expiration Date
Abstract
Disclosed are compositions and methods for directing proteins to specific loci in the genome and uses thereof. In one aspect, the disclosed methods allow for directing proteins to specific loci in the genome of an organism, including the steps of providing a fusion protein comprising a DNA localization component and an effector molecule. Preferred embodiments of the disclosure include, but are not limited to, the following fusion proteins: dSaCas9-Clo051, dCas9-Clo051, Xanthomonas-TALE-Clo051, and Ralstonia-TALE-Clo051.
Core Innovation
The described invention provides genome editing via an effector molecule that is a fusion protein. The effector includes an inactivated Cas9 (dCas9) or an inactivated nuclease domain, fused to Clo051 or a Clo051 nuclease domain, connected by a Gly-Gly-Gly-Gly-Ser peptide linker.
The disclosed system combines DNA localization with repeat-based DNA-sequence recognition and endonuclease activity. Clo051 is presented as a nuclease that functions independently of DNA binding and cleaves dsDNA, including dimer cleavage of dsDNA. Engineered endonuclease variants are described to preserve cleavage while disabling binding.
The disclosure further describes targeted genome modifications using homologous recombination to introduce additions/deletions in plants and other organisms. It also describes broader modular architectures for gene regulation and DNA modification, including modular repeat domains and transcription activators/repressors. Non-human organisms, including transgenic/non-human organisms, are described as embodiments for the disclosed modular effector concepts.
Claims Coverage
The independent claims identified in the provided claim set provide at least one core inventive arrangement: a composition that combines a DNA localization component with at least one gRNA and a fusion effector comprising dCas9 (or an inactivated nuclease domain) connected to Clo051 (or a Clo051 nuclease domain) by a Gly-Gly-Gly-Gly-Ser peptide linker.
DNA localization with gRNA and Clo051–dCas9 fusion connected by a Gly-Gly-Gly-Gly-Ser linker
A composition includes a DNA localization component comprising at least one guide RNA (gRNA) and an effector molecule that is a fusion protein with (i) an inactivated Cas9 (dCas9) or an inactivated nuclease domain thereof and (ii) Clo051 or a nuclease domain thereof, connected by a Gly-Gly-Gly-Gly-Ser peptide linker.
Based on the explicitly provided independent claim, the claimed coverage centers on a fusion effector architecture that links inactivated Cas9 (or an inactivated nuclease domain) to Clo051 (or a Clo051 nuclease domain) for use with a DNA localization component comprising at least one gRNA.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
Interested in licensing this patent?