This technology is an engineered CRISPR-associated transposon (CRISPR-CAST) platform that enables programmable, cut-free insertion of DNA into specific sites on the genome, allowing precise and efficient gene modification or payload integration in living cells.
Current approaches for inserting DNA into the genome rely on nuclease-induced double-strand breaks followed by cellular repair, a process that can be inefficient, unpredictable, and can lead to mutations or toxicity. These limitations are especially pronounced in mammalian cells, where double-strand breaks often yield low insertion rates and are difficult to control across the genome. Additionally, existing methods struggle to efficiently incorporate larger DNA fragments into the mammalian genome. Safer and more reliable strategies for targeted DNA integration are needed to reduce off-target effects and improve the efficiency of gene insertion in mammalian cells.
This technology is a platform that uses a CRISPR-associated transposon (CAST) approach to direct a DNA payload to a specific genomic site, enabling insertion without creating double-strand breaks. After the transposon machinery completes the integration reaction, it can remain stuck to the DNA, which can hinder the final steps of the insertion process in mammalian cells. To address this, the platform incorporates accessory factors such as ClpX unfoldase or engineered degron tags, which help remove the residual CAST transposomes, thereby allowing the cell’s own repair machinery to finalize the integration more efficiently.
This technology has been validated with mammalian cell lines.
Patent Pending (US 20250297289)
IR CU23140, CU23270
Licensing Contact: Cynthia Lang