[2PS-060]
우수논문발표상 응모자Development of a Lipid nanoparticle-Based DNA Delivery Platform for HDR-Mediated Genome Editing in Immune Cells
발표자최예림 (서울대학교)
연구책임자이혁진 (서울대학교)
Abstract
Non-viral genome editing systems supporting large DNA insertions are important for next-generation immune cell therapies. Although CRISPR/Cas9-mediated homology-directed repair (HDR) enables precise gene integration, its use is limited by inefficient donor DNA delivery and cytotoxicity associated with electroporation. To address these challenges, we developed a LNP platform optimized for delivery of large donor DNA templates for HDR knock-in. Comparative evaluation of plasmid DNA (pDNA) and linear double-stranded DNA (dsDNA) donors formulated with HHES ionizable lipids showed strong dependencies between donor format, formulation quality, and editing efficiency. dsDNA donors produced more uniform LNP populations and achieved higher HDR efficiencies than pDNA donors. HHES-based, all-in-one dsDNA HDR-LNPs achieved highest knock-in efficiencies. These findings expand LNP applications to enable delivery of large DNA donors for precise genome editing and support therapeutic genome editing.