Abstract
Efficient cytosolic delivery of nucleic acid therapeutics remains a central challenge for DNA nanostructure-based biomedical applications. Here, we report a programmable DNA origami delivery platformthe DNA Soccer Framework (DSF)cofunctionalized with fluorocarbon chains and sgc8 aptamers to enhance cellular uptake and promote endosomal escape for effective siRNA delivery. Fluorocarbon moieties of tunable lengths and densities were grafted onto DSF to facilitate lipid raft-mediated endocytosis, which favors cytosolic entry by bypassing lysosomal degradation. Concurrently, aptamer comodification enabled selective targeting of cancer cells. Systematic optimization revealed that a surface modification ratio of 3:1 (fluorocarbon to aptamer) among the 90 available sites yielded synergistic improvements in internalization and cytosolic release, significantly enhancing siRNA-mediated gene silencing. Upon loading with siRNA targeting Bcl2L12, the optimized DSF variant induced highest apoptosis in two cancer cell lines, while maintaining minimal cytotoxicity. This work establishes a robust and tunable platform for nucleic acid delivery, advancing the application of DNA nanotechnology in gene therapy and precision medicine.