Mass Production of Self-assembled Synthetic Virus Encapsulating Functional ssDNA in Living Bacteria, With Increased Activity and Resistance to Degradation in Human Blood Serum

The use of DNA nanotechnology tools in real-world applications requires bridging the gap between in vitro experiments and the rapid degradation of oligonucleotides(OGNS) under in vivo conditions. The major challenge is providing sufficient protection against various nucleases under “real-life” environments, such as body fluids, while maintaining the OGNS functionality and structure. Here, Escherichia coli was engineered to produce functional DNA nanostructures encapsulated inside an MS2 virus-like particles (VLPs), thus forming functional nanocomposites in vivo. This approach enhances the protection of the functional DNA structures against various enzymes, both in vivo and in situ, resulting in ssDNA stabilization. Furthermore, the encapsulated ssDNA lifetime is increased four-fold under the biological environment of concentrated blood serum. This system paves the way for bulk production of DNA nanostructures, and other in vivo applications such as programmed drug delivery and nanoreactors.