self-assembled synthetic virus enables large scale production of functional DNA, with increased activity and resistance to degradation in blood serum

Synthetic biology revolutionized the field of bioengineering, providing a transformative genetic language for the manipulation of cellular behaviors. Similarly, DNA nanotechnology prefects the engineering of molecular-scale devices with exquisite control over geometry and site-specific functionalization at the nanoscale. DNA nanotechnology has been extensively used for the development of drug delivery systems and biosensors. By integrating synthetic biology tools with DNA nanotechnology, we have recently resolved the challenge to encode and produce precise artificial DNA nanostructures in living bacteria. However, the major limitation of single-stranded DNA production in living bacteria is its rapid degradation in the presence of intra- and extra-cellular conditions and enzymes. Here, bacteria have been engineered to produce and encapsulate functional ssDNA in the form of DNA nanomachine and DNA aptamers inside MS2 virus-like particles (VLPs). This encapsulation protects the DNA structures, greatly enhancing their stability in situ. We further demonstrate the ability to encapsulate ssDNA from 200-nt to 1500-nt, while maintaining their structural functionality within the VLPs. We demonstrate this by encapsulating light-up DNA aptamer and DNA nanotweezers capable to sense and react to external changes. Moreover, The VLPs protect and increase the lifetime of the encapsulated ssDNA 4 fold in blood serum samples and increase the functional DNA activity within bacteria. This work paves the way for bulk production of DNA nanostructures, as well as the ability to be induced in cells for in vivo applications such as programmed drug delivery and nanoreactors.