Genetic encoding and expression of RNA origami cytoskeletons in synthetic cells [research data]

• Verfasst von: Tran, Mai P. [VerfasserIn]
• Verfasst von: Chakraborty, Taniya [VerfasserIn]
• Verfasst von: Poppleton, Erik [VerfasserIn]
• Verfasst von: Monari, Luca [VerfasserIn]
• Verfasst von: Illig, Maja [VerfasserIn]
• Verfasst von: Gießler, Franziska [VerfasserIn]
• Verfasst von: Göpfrich, Kerstin [VerfasserIn]
• Titel: Genetic encoding and expression of RNA origami cytoskeletons in synthetic cells [research data]
• Verf.angabe: Mai P. Tran, Taniya Chakraborty, Erik Poppleton, Luca Monari, Maja Illig, Franziska Giessler, Kerstin Göpfrich
• Verlagsort: Heidelberg
• Verlag: Universität
• E-Jahr: 2025
• Jahr: 2025-02-03
• Umfang: 1 Online-Ressource (8 Files)
• Fussnoten: Gefördert durch: Human Frontier Science Program (HFSP): RGP003/2023; European Research Council (ERC): ENSYNC (No. 101076997); Hector Fellow Academy; Federal Ministry of Education and Research (BMBF); Ministry of Science Baden-Württemberg: Excellence Strategy ; Gesehen am 05.02.2025
• DOI: doi:10.11588/DATA/LJLAMX
• Datenträger: Online-Ressource
• Dokumenttyp: Forschungsdaten
• Dokumenttyp: Datenbank
• Sprache: eng
• Sonstige Nr.: Grant number: HFSP RGP003/2023
• Sonstige Nr.: Grant number: ENSYNC 101076997
• Sach-SW: Chemistry
• Sach-SW: Engineering
• Sach-SW: Health and Life Sciences
• Sach-SW: Medicine
• Sach-SW: Physics
• K10plus-PPN: 1916388817
• K10plus-PPN:
  Universitätsbibliothek
• Lokale URL UB: Zum Volltext

Abstract

Bottom-up synthetic biology seeks to engineer a cell from molecular building blocks. Using DNA nanotechnology, building blocks, such as cytoskeletons, have been reverse-engineered. However, DNA nanostructures rely on chemical synthesis and thermal annealing, and therefore synthetic cells cannot produce them from their constituents such as nucleotides. Here, we introduce RNA origami cytoskeleton mimics as alternative nucleic acid-based molecular hardware for synthetic cells, which we express directly inside GUVs containing a DNA template and a polymerase, chemically fuelled by feeding nucleotides from the outside. We designed RNA origami tiles that fold upon transcription and self-assemble into micrometer-long, 3D RNA origami nanotubes under isothermal conditions. We observe sequence mutations on the DNA template lead to RNA origami nanotubes and closed-ring phenotypes. Molecular dynamics simulations show that these phenotypic transitions are governed by alterations in the stability of RNA secondary structures. Additionally, we achieve cortex formation with aptamer-functionalized RNA nanotubes and show that nanotube polymerization leads to membrane deformation. Altogether, our data suggest that the expression of RNA origami-based hardware will help to explore active, evolvable, and RNA-based synthetic cells.