Designer RNA nanostructures co-transcribed and self-assembled inside human cell nuclei

Area of Science:

• Synthetic Biology
• Molecular Biology
• Nanotechnology

Background:

• Interfacing with and regulating cellular processes using nucleic acid nanostructures is challenging.
• Nuclear delivery and retention of synthetic nanostructures in eukaryotic cells are significant hurdles.

Purpose of the Study:

• To present a platform for genetically encoded, self-assembling RNA nanostructures.
• To demonstrate their co-transcriptional production, nuclear assembly, and functional integration within live human cells.

Main Methods:

• Co-transcriptional folding of single-stranded RNAs into defined nanostructures (rings, ribbons, nanonets).
• In vitro validation using atomic force microscopy.
• Functional integration of fluorescent aptamers and RNA sensing capabilities.
• In vivo demonstration in live human cells using confocal live-cell imaging and transmission electron microscopy.

Main Results:

• Formation of RNA nanostructures with programmable geometry (rings, ribbons, nanonets) validated in vitro.
• Successful co-transcriptional production and assembly of RNA nanonets within the nucleus of live human cells.
• Demonstrated retention of well-defined nanostructure patterns in the nucleus.
• Functional integration of aptamers and sensing capabilities within the nanostructures.

Conclusions:

• Established a genetically encoded, self-assembling RNA nanostructure system.
• Demonstrated programmable geometry and nuclear localization capabilities.
• Provided a foundation for RNA-based nanodevices for studying biological properties in live cells and tissues.