Extrusion of RNA from a DNA-Origami-Based Nanofactory

• Published in: ACS nano 2020-02, Vol.14 (2), p.1550-1559
• Main Authors: Hahn, Jaeseung, Chou, Leo Y.T, Sørensen, Rasmus S, Guerra, Richard M, Shih, William M
• Format: Article
• Language: English
• Subjects: DNA - chemistry; DNA - metabolism; DNA-Directed RNA Polymerases - metabolism; Endoribonucleases - metabolism; Nanotechnology; Particle Size; RNA - biosynthesis; RNA - chemistry; Surface Properties
• ISSN: 1936-0851; 1936-086X
• DOI: 10.1021/acsnano.9b06466

Cells often spatially organize biomolecules to regulate biological interactions. Synthetic mimicry of complex spatial organization may provide a route to similar levels of control for artificial systems. As a proof-of-principle, we constructed an RNA-extruding nanofactory using a DNA-origami barrel with an outer diameter of 60 nm as a chassis for integrated rolling-circle transcription and processing of RNA through spatial organization of DNA templates, RNA polymerases, and RNA endonucleases. The incorporation efficiency of molecular components was quantified to be roughly 50% on designed sites within the DNA-origami chassis. Each integrated nanofactory with RNA-producing units, composed of DNA templates and RNA polymerases, produced 100 copies of target RNA in 30 min on average. Further integration of RNA endonucleases that cleave rolling-circle transcripts from concatemers into monomers resulted in 30% processing efficiency. Disabling spatial organization of molecular components on DNA origami resulted in suppression of RNA production as well as processing.