NUCLEOBASE EDITORS AND USES THEREOF

Smart overview of the Invention

Strategies and systems for targeted nucleic acid editing are detailed, focusing on editing specific genome sites, like those in humans. Fusion proteins combining Cas9 with nucleic acid editing proteins, such as deaminase domains, are central to these methods. The application emphasizes the creation of reagents and kits to facilitate the generation of these fusion proteins for precise genome editing.

Background

The CRISPR-Cas9 system, a prokaryotic adaptive immune mechanism, has been adapted for genome engineering. Cas9, guided by RNA molecules, targets specific DNA sequences for editing. The dead Cas9 (dCas9) variant retains DNA binding capabilities without nuclease activity, enabling it to target various proteins to specific DNA sites for applications beyond nucleases.

Innovation

This application introduces fusion proteins comprising dCas9 and deaminase domains for efficient nucleobase editing. These fusion proteins are engineered to deaminate target cytidine residues with high precision and efficiency. The use of specific linkers enhances the performance of these base editors by reducing unwanted mutations and increasing target specificity.

Applications

The disclosed technology offers significant potential in gene therapy and research by enabling precise single-base modifications in genomic DNA. This could address genetic disorders caused by single nucleotide changes. The fusion proteins can be tailored into various genome engineering tools, including transcriptional activators and repressors, enhancing their utility in genetic research and therapeutic applications.

Technical Details

• Fusion Proteins: Composed of dCas9 and deaminase domains, such as APOBEC1.
• Linkers: Specific amino acid sequences like SGSETPGTSESATPES improve efficiency.
• Deaminases: Include APOBEC family members from various organisms.
• Editing Precision: Minimizes off-target effects while maximizing desired modifications.