Temperature-Controlled Encapsulation and Release of an Active Enzyme in the Cavity of a Self-Assembled DNA Nanocage

Juul, Sissel; Iacovelli, Federico; Falconi, Mattia; Kragh, Sofie L.; Christensen, Brian; Frøhlich, Rikke; Franch, Oskar; Kristoffersen, Emil L.; Stougaard, Magnus; Leong, Kam W.; Ho, Yi‐Ping; Sørensen, Esben S.; Birkedal, Victoria; Desideri, Alessandro; Knudsen, Birgitta R.

doi:10.1021/nn4030543

Cited by 142 publications

(166 citation statements)

References 74 publications

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“…9,21 Three-dimensional (3D) RNA nanostructures can also be used as scaffolding to direct high precision assembly of nano-sized materials, such as colloidal particles and membrane proteins, into objects with the desired spacing, shape and organisation. 1,22,23 …”

Section: Introductionmentioning

confidence: 99%

Assembly of RNA nanostructures on supported lipid bilayers

et al. 2015

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The assembly of nucleic acid nanostructures with controlled size and shape has large impact in the fields of nanotechnology, nanomedicine and synthetic biology. The directed arrangement of nanostructures at interfaces is important for many applications. In spite of this, the use of laterally mobile lipid bilayers to control RNA three-dimensional nanostructure formation on surfaces remains largely unexplored. Here, we direct the self-assembly of RNA building blocks into three-dimensional structures of RNA on fluid lipid bilayers composed of cationic 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP) or mixtures of zwitterionic 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC) and cationic sphingosine. We demonstrate the stepwise supramolecular assembly of discrete building blocks through specific and selective RNA-RNA interactions, based on results from quartz crystal microbalance with dissipation (QCM-D), ellipsometry, fluorescence recovery after photobleaching (FRAP) and total internal reflection fluorescence microscopy (TIRF) experiments. The assembly can be controlled to give a densely packed single layer of RNA polyhedrons at the fluid lipid bilayer surface. We show that assembly of the 3D structure can be modulated by sequence specific interactions, surface charge and changes in the salt composition and concentration. In addition, the tertiary structure of the RNA polyhedron can be controllably switched from an extended structure to one that is dense and compact. The versatile approach to building up three-dimensional structures of RNA does not require modification of the surface or the RNA molecules, and can be used as a bottom-up means of nanofabrication of functionalized bio-mimicking surfaces.

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Section: Introductionmentioning

confidence: 99%

Assembly of RNA nanostructures on supported lipid bilayers

et al. 2015

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“…Through the DNA selfassembly process, DNA molecules could form various cage structures, including tetrahedra, cubes, octahedra, dodecahedra and icosahedra shapes. Juul et al [53] conducted experiments to control the encapsulation and release of horseradish peroxidase in a DNA nanocavity through temperature change. The nanocavity (approximately 10-12 nm) was covalently closed by 12 double-stranded DNA helices as the edges of the structure.…”

Section: Nanocontainersmentioning

confidence: 99%

Nanobiocatalyst advancements and bioprocessing applications

Misson

Zhang

Jin

2015

J. R. Soc. Interface.

214

108

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The nanobiocatalyst (NBC) is an emerging innovation that synergistically integrates advanced nanotechnology with biotechnology and promises exciting advantages for improving enzyme activity, stability, capability and engineering performances in bioprocessing applications. NBCs are fabricated by immobilizing enzymes with functional nanomaterials as enzyme carriers or containers. In this paper, we review the recent developments of novel nanocarriers/nanocontainers with advanced hierarchical porous structures for retaining enzymes, such as nanofibres (NFs), mesoporous nanocarriers and nanocages. Strategies for immobilizing enzymes onto nanocarriers made from polymers, silicas, carbons and metals by physical adsorption, covalent binding, cross-linking or specific ligand spacers are discussed. The resulting NBCs are critically evaluated in terms of their bioprocessing performances. Excellent performances are demonstrated through enhanced NBC catalytic activity and stability due to conformational changes upon immobilization and localized nanoenvironments, and NBC reutilization by assembling magnetic nanoparticles into NBCs to defray the high operational costs associated with enzyme production and nanocarrier synthesis. We also highlight several challenges associated with the NBC-driven bioprocess applications, including the maturation of large-scale nanocarrier synthesis, design and development of bioreactors to accommodate NBCs, and long-term operations of NBCs. We suggest these challenges are to be addressed through joint collaboration of chemists, engineers and material scientists. Finally, we have demonstrated the great potential of NBCs in manufacturing bioprocesses in the near future through successful laboratory trials of NBCs in carbohydrate hydrolysis, biofuel production and biotransformation.

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“…Juul et al [62] developed a preassembled DNA cage that can encapsulate and release enzymes by temperature-induced conformational changes in the assembly (illustrated in Figure 5e). The cage-like structure was assembled using 12 DNA duplexes that formed the edges of the design.…”

Section: Enzyme Containers and Carriersmentioning

confidence: 99%

“…The closed box efficiently protects the assembled enzyme cascade pair from protease digestion [59]; ( c ) A DNA origami-based nanocarrier loaded with luciferase (LUC) enzymes. The enzyme activity can be tuned by coating the carrier with cationic polymers [60]; ( d ) A light-triggered release of proteins such as bovine serum albumin (BSA) from DNA origami container [61]; ( e ) A DNA cage that can trap and release an enzyme (HRP) through the temperature-controlled conformational changes [62]; ( f ) β-galactosidase (β-gal) can be coated by DNA strands for significantly enhanced cellular delivery [63]. A box with a lid in ( a ) is reproduced with permission from [56].…”

Section: Figurementioning

confidence: 99%

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DNA-Based Enzyme Reactors and Systems

et al. 2016

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During recent years, the possibility to create custom biocompatible nanoshapes using DNA as a building material has rapidly emerged. Further, these rationally designed DNA structures could be exploited in positioning pivotal molecules, such as enzymes, with nanometer-level precision. This feature could be used in the fabrication of artificial biochemical machinery that is able to mimic the complex reactions found in living cells. Currently, DNA-enzyme hybrids can be used to control (multi-enzyme) cascade reactions and to regulate the enzyme functions and the reaction pathways. Moreover, sophisticated DNA structures can be utilized in encapsulating active enzymes and delivering the molecular cargo into cells. In this review, we focus on the latest enzyme systems based on novel DNA nanostructures: enzyme reactors, regulatory devices and carriers that can find uses in various biotechnological and nanomedical applications.

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Temperature-Controlled Encapsulation and Release of an Active Enzyme in the Cavity of a Self-Assembled DNA Nanocage

Cited by 142 publications

References 74 publications

Assembly of RNA nanostructures on supported lipid bilayers

Assembly of RNA nanostructures on supported lipid bilayers

Nanobiocatalyst advancements and bioprocessing applications

DNA-Based Enzyme Reactors and Systems

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