METHOD FOR PRODUCING DNA-EDITED EUKARYOTIC CELL, AND KIT USED IN THE SAME

Smart overview of the Invention

A novel method for producing DNA-edited eukaryotic cells has been developed using the CRISPR-Cas3 system. Unlike the widely utilized CRISPR-Cas9 system, which has been effective in various eukaryotic cells, the CRISPR-Cas3 system previously faced challenges in achieving genome editing in these cells. The method leverages the unique properties of the CRISPR-Cas3 system to enhance editing efficiency and applicability across different types of eukaryotic organisms.

Technical Background

The CRISPR-Cas systems are adaptive immune mechanisms found in bacteria and archaea, which protect against foreign genetic material. These systems can be classified into Class 1 and Class 2, with Class 1 including type I systems that utilize Cas3 proteins. Prior attempts to implement CRISPR-Cas3 systems for genomic editing in eukaryotic cells had not yielded successful results, leading to a focus on enhancing their functionality for this purpose.

Methodology for DNA Editing

The method involves introducing a CRISPR-Cas3 system into eukaryotic cells, which consists of a Cas3 protein, a Cascade protein, and a crRNA. Notably, the use of pre-crRNA instead of mature crRNA is critical for efficient genomic editing. Additionally, incorporating a nuclear localization signal into the Cas3 protein further boosts editing efficiency, allowing for significant deletions within target DNA sequences.

Applications Beyond Eukaryotic Cells

This innovative approach is not limited to just eukaryotic cells; it can also be applied to animals (excluding humans) and plants. The versatility of the CRISPR-Cas3 system opens up new possibilities for genetic modifications across various biological systems, potentially enhancing agricultural practices and biomedical research.

Advantages and Future Implications

The establishment of an effective CRISPR-Cas3 system in eukaryotic cells marks a significant advancement in genome editing technology. The ability to edit DNA with greater precision and efficiency could lead to breakthroughs in gene therapy, crop improvement, and synthetic biology. This method represents a promising tool for researchers aiming to explore genetic interventions more effectively than ever before.