3 research outputs found
Composing RNA Nanostructures from a Syntax of RNA Structural Modules
Natural
stable RNAs fold and assemble into complex three-dimensional
architectures by relying on the hierarchical formation of intricate,
recurrent networks of noncovalent tertiary interactions. These sequence-dependent
networks specify RNA structural modules enabling orientational and
topological control of helical struts to form larger self-folding
domains. Borrowing concepts from linguistics, we defined an extended
structural syntax of RNA modules for programming RNA strands to assemble
into complex, responsive nanostructures under both thermodynamic and
kinetic control. Based on this syntax, various RNA building blocks
promote the multimolecular assembly of objects with well-defined three-dimensional
shapes as well as the isothermal folding of long RNAs into complex
single-stranded nanostructures during transcription. This work offers
a glimpse of the limitless potential of RNA as an informational medium
for designing programmable and functional nanomaterials useful for
synthetic biology, nanomedicine, and nanotechnology
Fluorescent Monitoring of RNA Assembly and Processing Using the Split-Spinach Aptamer
As
insights into RNA’s many diverse cellular roles continue
to be gained, interest and applications in RNA self-assembly and dynamics
remain at the forefront of structural biology. The bifurcation of
functional molecules into nonfunctional fragments provides a useful
strategy for controlling and monitoring cellular RNA processes and
functionalities. Herein we present the bifurcation of the preexisting
Spinach aptamer and demonstrate its utility as a novel split aptamer
system for monitoring RNA self-assembly as well as the processing
of pre-short interfering substrates. We show for the first time that
the Spinach aptamer can be divided into two nonfunctional halves that,
once assembled, restore the original fluorescent signal characteristic
of the unabridged aptamer. In this regard, the split-Spinach aptamer
is represented as a potential tool for monitoring the self-assembly
of artificial and/or natural RNAs
Co-transcriptional Assembly of Chemically Modified RNA Nanoparticles Functionalized with siRNAs
We report a generalized methodology for the one-pot production
of chemically modified functional RNA nanoparticles during in vitro
transcription with T7 RNA polymerase. The efficiency of incorporation
of 2′-fluoro-dNTP in the transcripts by the wild type T7 RNA
polymerase dramatically increases in the presence of manganese ions,
resulting in a high-yield production of chemically modified RNA nanoparticles
functionalized with siRNAs that are resistant to nucleases from human
blood serum. Moreover, the unpurified transcription mixture can be
used for functional ex vivo pilot experiments