ABSTRACT
Both DNA- and RNA-based nanotechnologies are remarkably useful for in vitro molecular-scale device engineering and are applied in a vast array of applications. However, while the function of nucleic acid nanostructures is robust under in vitro settings, their implementation in real-world conditions requires overcoming their inherent degradation sensitivity and subsequent loss of function. Viruses are minimalistic yet sophisticated supramolecular assemblies, able to protect their nucleic acid content in inhospitable biological environments. Inspired by this natural ability, we engineered RNA-virus-like particles (VLPs) nanocarriers (NCs). We showed that the VLPs can serve as an excellent protective shell against nuclease-mediated degradation. We then harnessed biological recognition elements and demonstrated how engineered riboswitch NCs can act as a possible disease-modifying treatment for genetic metabolic disorders. The functional riboswitch is capable of selectively and specifically binding metabolites and preventing their self-assembly process and its downstream effects. When applying the riboswitch nano-carriers to an in vivo yeast model of adenine accumulation and self-assembly, significant inhibition of the sensitivity to adenine feeding was observed. In addition, using an amyloid-specific dye, we proved the riboswitch nano-carriers ability to reduce the level of intracellular amyloid-like cytotoxic structures. The potential of this RNA therapeutic technology does not stop at metabolic disorders, as it can be easily fine-tuned to be applied to other conditions and diseases.
Competing Interest Statement
S.Z.T, D.G and E.G has filed for a provisional patent application for the described technology. USP No. 63/249,834 filed 29/09/2021 and 63/337,734 filed 3 May 2022
ABBREVIATIONS
- WT
- wild type
- TR-RNA
- translational repression RNA
- VLPs
- virus like particles
- APRT
- adenine phosphoribosyltransferase
- ADA
- adenosine deaminase
- IEM
- inborn error of metabolism
- TEM
- transmission electron microscopy
- NCs
- nanocarriers
- TA
- Tannic Acid