Angle-controllable RNA tiles for programable array assembly and RNA sensing.

Area of Science:

• Biomaterials Science
• Synthetic Biology
• RNA Nanotechnology

Background:

• Programmed self-assembly of RNA nanostructures is key for tailored biomaterials.
• Current engineered RNA nanostructures have limitations in variety, complexity, and programmability.

Purpose of the Study:

• To introduce a new category of artificially designed RNA tiles with controllable angles (65° or 90°).
• To expand the collection of engineered multi-stranded RNA tiles.
• To explore design strategies for improved array assembly and functionalization.

Main Methods:

• Integration of antiparallel crossovers and T-junctions to create novel RNA tiles.
• Investigation of T-loop configuration, sticky end pairing, and annealing methods for assembly.
• Co-transcriptional folding of a single-stranded RNA tile.
• Incorporation of split broccoli RNA aptamers for fluorescence activation.

Main Results:

• 22 distinct multi-stranded RNA tiles were designed, significantly increasing available options.
• Design strategies influencing array assembly were identified.
• A single-stranded RNA tile capable of co-transcriptional folding was developed.
• Programmable fluorescence activation along linear RNA arrays was demonstrated using split broccoli aptamers.

Conclusions:

• The new RNA tiles offer enhanced programmability and structural control for nanostructure assembly.
• These findings expand the toolkit for creating complex RNA nanostructures for biomaterial applications.
• The integration of functional elements like aptamers opens avenues for programmable sensing and molecular engineering.