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. 2016 Sep;28(34):7501-7.
doi: 10.1002/adma.201601976. Epub 2016 Jun 20.

Controllable Self-Assembly of RNA Tetrahedrons With Precise Shape and Size for Cancer Targeting

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Free PMC article

Controllable Self-Assembly of RNA Tetrahedrons With Precise Shape and Size for Cancer Targeting

Hui Li et al. Adv Mater. .
Free PMC article

Abstract

RNA tetrahedral nanoparticles with two different sizes are successfully assembled by a one-pot bottom-up approach with high efficiency and thermal stability. The reported design principles can be extended to construct higher-order polyhedral RNA architectures for various applications such as targeted cancer imaging and therapy.

Keywords: Phi29 packaging RNA; RNA nanoparticles; RNA nanotechnology; tetrahedrons; three-way junctions.

Figures

Figure 1
Figure 1. Design and assembly of 8 nm RNA tetrahedrons
(a) 2D sequence of pRNA monomer showing the central pRNA-3WJ motif. The 22-nucleotide core sequence (with red color) of pRNA-3WJ are used to construct RNA tetrahedrons. (b) 2D sequences and (c) 3D computational model of RNA tetrahedrons. (d) 7% native PAGE gel showing step-wise assembly of RNA tetrahedrons. ‘+’ indicates the presence of the strands. M: ultra low range DNA Ladder. (e) AFM images and (f) Single particle cryo-EM 3D reconstruction of 8 nm RNA tetrahedrons.
Figure 2
Figure 2. Physiochemical characterization of RNA tetrahedrons
Dynamic Light Scattering (DLS) assay showing (a) the hydrodynamic size and (b) the zeta potential of RNA tetrahedrons. (c) Melting curve of RNA tetrahedron complex and each of the four component strands. (d) Comparison of melting curves for RNA, 2′-F and DNA tetrahedrons.
Figure 3
Figure 3. Design, assembly and characterization of 17 nm RNA tetrahedrons
(a) Schematic showing tunable size conversion (from 22 bp per edge to 55 bp per edge) of RNA tetrahedrons. (b) 6% native PAGE gel showing step-wise assembly of larger RNA tetrahedrons. ‘+’ indicates the presence of the strands. M: 100bp DNA ladder. (c) DLS assay showing the hydrodynamic size of larger RNA tetrahedrons. (d) AFM images and (e) Cryo-EM images and 3D reconstruction of RNA tetrahedrons.
Figure 4
Figure 4. Functional characterization of multifunctional RNA tetrahedrons
(a) Schematic showing multifunctional RNA tetrahedrons harboring HBV ribozyme, MG aptamer, Spinach aptamer and STV aptamer. (b) 7% native PAGE gel showing step-wise assembly of multifunctional RNA tetrahedrons. ‘+’ indicates the presence of the strands. (c) Ribozyme activity assay showing cleavage of 135 nt substrate. Fluorogemic assay demonstrating fluorescence emission of (d) MG aptamer and (e) Spinach aptamer. (f) Streptavidin (STV) aptamer binding assay using STV affinity column.
Figure 5
Figure 5. In vitro and in vivo evaluation of RNA tetrahedrons harboring siRNA and cancer-targeting aptamers
(a) Confocal images showing RNA tetrahedron (with and without EGFR aptamers) binding to MDA-MB-231 cells. (b) Luciferase siRNA silencing effects assayed by dual luciferase assay. Error bars indicate mean ± SD. (c) Biodistribution assay in orthotopic MDA-MB-231 tumor-bearing mice after systemic tail vein injection of RNA tetrahedrons harboring EGFR aptamers.

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