Engineered Riboswitch Nanocarriers as a Possible Disease-Modifying Treatment for Metabolic Disorders

Abstract

Both DNA- and RNA-based nanotechnologies are remarkably useful for the engineering of molecular devices in vitro and are applied in a vast collection of applications. Yet, the ability to integrate functional nucleic acid nanostructures in applications outside of the lab requires overcoming their inherent degradation sensitivity and subsequent loss of function. Viruses are minimalistic yet sophisticated supramolecular assemblies, capable of shielding their nucleic acid content in nuclease-rich environments. Inspired by this natural ability, we engineered RNA-virus-like particles (VLPs) nanocarriers (NCs). We showed that the VLPs can function as an exceptional 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 nanocarriers 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 nanocarriers’ ability to reduce the level of intracellular amyloid-like metabolite cytotoxic structures. The potential of this RNA therapeutic technology does not apply only to metabolic disorders, as it can be easily fine-tuned to be applied to other conditions and diseases.

Molecular Complementarity of Proteomimetic Materials for Target‐Specific Recognition and Recognition‐Mediated Complex Functions

“…[276][277][278] Catalytic riboswitches have been also included in mammalian cells as transient induction systems for clinical use. [224,279,280] By sharing the common mechanism of gene expression regulation with riboswitches, toehold switches that use oligonucleotides as input targets have served as updated RNA-encoded biosensors. [213] Based on simple Watson-Crick base pairing, their structure can be readily reconstructed to recognize the different sequences of nucleic acids, and the change in gene expression is typically significantly larger than that of the riboswitches (Figure 4d).…”

“…[ 276–278 ] Catalytic riboswitches have been also included in mammalian cells as transient induction systems for clinical use. [ 224,279,280 ]…”

Magnetically Induced Thermal Effects on Tobacco Mosaic Virus-Based Nanocomposites for a Programmed Disassembly of Protein Cages

“…Among these innovative protein-based nanosystems, viruses stand out as nanostructures with precisely defined shapes designed to protect and transport their genetic payload. These viral protein cages have demonstrated efficacy as cargo carriers, , anticancer therapeutics, , and vaccines. − Moreover, native virus capsids possess unique assembly properties, rendering them attractive building blocks for bottom-up nanobioengineering solutions, particularly when combined with DNA. , In particular, nanoparticles derived from tobacco mosaic virus (TMV) and cowpea chlorotic mottle virus (CCMV) have emerged as promising carriers for targeted drug delivery due to their nonpathogenic nature in humans and scalability in production. − Plant viruses have also been investigated as gene delivery vectors, given that they lack the machinery necessary to navigate mammalian cells efficiently, hindering therefore trafficking, unpacking, and gene expression.…”

“…Among these innovative protein-based nanosystems, viruses stand out as nanostructures with precisely defined shapes designed to protect and transport their genetic payload. These viral protein cages have demonstrated efficacy as cargo carriers, 3 , 4 anticancer therapeutics, 5 , 6 and vaccines. 7 − 9 Moreover, native virus capsids possess unique assembly properties, rendering them attractive building blocks for bottom-up nanobioengineering solutions, particularly when combined with DNA.…”