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TitleStructure, folding and flexibility of co-transcriptional RNA origami.
Journal, issue, pagesNat Nanotechnol, Vol. 18, Issue 7, Page 808-817, Year 2023
Publish dateFeb 27, 2023
AuthorsEwan K S McRae / Helena Østergaard Rasmussen / Jianfang Liu / Andreas Bøggild / Michael T A Nguyen / Nestor Sampedro Vallina / Thomas Boesen / Jan Skov Pedersen / Gang Ren / Cody Geary / Ebbe Sloth Andersen /
PubMed AbstractRNA origami is a method for designing RNA nanostructures that can self-assemble through co-transcriptional folding with applications in nanomedicine and synthetic biology. However, to advance the ...RNA origami is a method for designing RNA nanostructures that can self-assemble through co-transcriptional folding with applications in nanomedicine and synthetic biology. However, to advance the method further, an improved understanding of RNA structural properties and folding principles is required. Here we use cryogenic electron microscopy to study RNA origami sheets and bundles at sub-nanometre resolution revealing structural parameters of kissing-loop and crossover motifs, which are used to improve designs. In RNA bundle designs, we discover a kinetic folding trap that forms during folding and is only released after 10 h. Exploration of the conformational landscape of several RNA designs reveal the flexibility of helices and structural motifs. Finally, sheets and bundles are combined to construct a multidomain satellite shape, which is characterized by individual-particle cryo-electron tomography to reveal the domain flexibility. Together, the study provides a structural basis for future improvements to the design cycle of genetically encoded RNA nanodevices.
External linksNat Nanotechnol / PubMed:36849548 / PubMed Central
MethodsEM (single particle) / EM (tomography)
Resolution4.08 - 27.0 Å
Structure data

EMDB-13592: Cryo-EM SPA reconstruction of an extended RNA origami 5 helix tile (5HT-B-3X)
Method: EM (single particle) / Resolution: 6.5 Å

EMDB-13625: Mature conformer 2 of a 6-helix bundle of RNA with a clasp
Method: EM (single particle) / Resolution: 7.43 Å

EMDB-13626: Young conformer 2 of a 6-helix bundle of RNA with a clasp
Method: EM (single particle) / Resolution: 7.04 Å

EMDB-13627: 6-Helix bundle of RNA
Method: EM (single particle) / Resolution: 6.61 Å

EMDB-13628, PDB-7ptk:
Young conformer of a 6-helix bundle of RNA with clasp
Method: EM (single particle) / Resolution: 5.18 Å

EMDB-13630, PDB-7ptl:
Mature conformer of a 6-helix bundle of RNA with clasp
Method: EM (single particle) / Resolution: 4.9 Å

EMDB-13633, PDB-7ptq:
RNA origami 5-helix tile
Method: EM (single particle) / Resolution: 4.08 Å

EMDB-13636, PDB-7pts:
RNA origami 5-helix tile
Method: EM (single particle) / Resolution: 5.71 Å

EMDB-13926, PDB-7qdu:
Twist-corrected RNA origami 5-helix Tile A
Method: EM (single particle) / Resolution: 5.14 Å

EMDB-28669: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 01)
Method: EM (tomography) / Resolution: 27.0 Å

EMDB-28670: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 02)
Method: EM (tomography) / Resolution: 25.0 Å

EMDB-28671: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 03)
Method: EM (tomography) / Resolution: 25.0 Å

EMDB-28672: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 04)
Method: EM (tomography) / Resolution: 24.0 Å

EMDB-28673: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 05)
Method: EM (tomography) / Resolution: 26.0 Å

EMDB-28674: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 06)
Method: EM (tomography) / Resolution: 27.0 Å

EMDB-28675: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 07)
Method: EM (tomography) / Resolution: 26.0 Å

EMDB-28676: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 08)
Method: EM (tomography) / Resolution: 25.0 Å

EMDB-28677: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 09)
Method: EM (tomography) / Resolution: 25.0 Å

EMDB-28678: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 10)
Method: EM (tomography) / Resolution: 26.0 Å

EMDB-28679: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 11)
Method: EM (tomography) / Resolution: 23.0 Å

EMDB-28680: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 12)
Method: EM (tomography) / Resolution: 23.0 Å

EMDB-28681: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 13)
Method: EM (tomography) / Resolution: 24.0 Å

EMDB-28682: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 14)
Method: EM (tomography) / Resolution: 27.0 Å

EMDB-28683: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 15)
Method: EM (tomography) / Resolution: 26.0 Å

EMDB-28684: Single-Molecule 3D Image of 16 Helix RNA Origami Satellite by Individual Particle Electron Tomography (No. 16)
Method: EM (tomography) / Resolution: 23.0 Å

Source
  • synthetic construct (others)
  • unidentified (others)
KeywordsRNA / origami / nanostructure

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