期刊
ACS NANO
卷 14, 期 2, 页码 1550-1559出版社
AMER CHEMICAL SOC
DOI: 10.1021/acsnano.9b06466
关键词
DNA nanotechnology; DNA origami; T7 RNA polymerase; Cas6e ribonuclease; catenane DNA; nanofactory; rolling-circle transcription
类别
资金
- Banting Fellowship
- NSERC
- National Science Foundation Graduate Research Fellowship [1122374]
- Carlsberg Foundation [2013_01_0736, CF14-0858]
- NSF Expeditions [CCF-1317291]
- Army Research Office [W911NF-12-1-0420]
- Institute for RNA Medicine (BIDMC) Pilot Award
- Wyss Institute at Harvard Core Faculty Award
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.
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