Optimizing ssDNA nanostructures production in vivo

It has been long known that single-stranded DNA (ssDNA) can be utilized to engineer sophisticated self-assembled nanostructures for medical and biotechnological applications. We have previously shown, for the first time, the engineering and production of ssDNA in vivo by harnessing the biological machinery of reverse transcription. This process requires an RNA structure primer for the reverse transcriptase enzyme (HTBS).

Here we show the ability to further optimize our system by removing the HTBS RNA structure in vivo using a self-cleaving DNAzyme, yielding a more precise in vivo product. Following cleavage, the ssDNAs maintain their functionality by forming DNA light-up aptamers, functional DNA structures which also serve as the ssDNA production sensors. Thus, we were able to detect ssDNA in vivo without its extraction, leading us towards developing DNA sensors and other biotechnological applications.

By advancing our ability to build ssDNA nanostructures in vivo we hope to accelerate the research of functional DNA applications such as: drug delivery systems, bio-sensors, intercellular reporters, gene regulators and more.