| 1. |
- Rahman, Mohammad A., et al.
(författare)
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Regeneration of Assembled, Molecular-Motor-Based Bionanodevices
- 2019
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Ingår i: Nano Letters. - : American Chemical Society (ACS). - 1530-6984 .- 1530-6992. ; 19:10, s. 7155-7163
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Tidskriftsartikel (refereegranskat)abstract
- The guided gliding of cytoskeletal filaments, driven by biomolecular motors on nano/microstructured chips, enables novel applications in biosensing and biocomputation. However, expensive and time-consuming chip production hampers the developments. It is therefore important to establish protocols to regenerate the chips, preferably without the need to dismantle the assembled microfluidic devices which contain the structured chips. We here describe a novel method toward this end. Specifically, we use the small, nonselective proteolytic enzyme, proteinase K to cleave all surface-adsorbed proteins, including myosin and kinesin motors. Subsequently, we apply a detergent (5% SDS or 0.05% Triton X100) to remove the protein remnants. After this procedure, fresh motor proteins and filaments can be added for new experiments. Both, silanized glass surfaces for actin-myosin motility and pure glass surfaces for microtubule-kinesin motility were repeatedly regenerated using this approach. Moreover, we demonstrate the applicability of the method for the regeneration of nano/microstructured silicon-based chips with selectively functionalized areas for supporting or suppressing gliding motility for both motor systems. The results substantiate the versatility and a promising broad use of the method for regenerating a wide range of protein-based nano/microdevices.
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| 2. |
- Reuther, Cordula, et al.
(författare)
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Comparison of actin- and microtubule-based motility systems for application in functional nanodevices
- 2021
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Ingår i: New Journal of Physics. - : IOP Publishing. - 1367-2630. ; 23:7
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Tidskriftsartikel (refereegranskat)abstract
- Over the last 25 years, extensive progress has been made in developing a range of nanotechnological applications where cytoskeletal filaments and molecular motors are key elements. This includes novel, highly miniaturized lab on a chip systems for biosensing, nanoseparation etc but also new materials and parallel computation devices for solving otherwise intractable mathematical problems. For such approaches, both actin-based and microtubule-based cytoskeletal systems have been used. However, in accordance with their different cellular functions, actin filaments and microtubules have different properties and interaction kinetics with molecular motors. Therefore, the two systems obviously exhibit different advantages and encounter different challenges when exploited for applications. Specifically, the achievable filament velocities, the capability to guide filaments along nanopatterned tracks and the capability to attach and transport cargo differ between actin- and microtubule-based systems. Our aim here is to systematically elucidate these differences to facilitate design of new devices and optimize future developments. We first review the cellular functions and the fundamental physical and biochemical properties of actin filaments and microtubules. In this context we also consider their interaction with molecular motors and other regulatory proteins that are of relevance for applications. We then relate these properties to the advantages and challenges associated with the use of each of the motor-filament systems for different tasks. Finally, fundamental properties are considered in relation to some of the most interesting future development paths e.g. in biosensing and biocomputation.
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| 3. |
- Reuther, Cordula, et al.
(författare)
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Molecular motor-driven filament transport across three-dimensional, polymeric micro-junctions
- 2021
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Ingår i: New Journal of Physics. - : Institute of Physics Publishing (IOPP). - 1367-2630. ; 23:12
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Tidskriftsartikel (refereegranskat)abstract
- Molecular motor-driven filament systems have been extensively explored for biomedical and nanotechnological applications such as lab-on-chip molecular detection or network-based biocomputation. In these applications, filament transport conventionally occurs in two dimensions (2D), often guided along open, topographically and/or chemically structured channels which are coated by molecular motors. However, at crossing points of different channels the filament direction is less well determined and, though crucial to many applications, reliable guiding across the junction can often not be guaranteed. We here present a three-dimensional (3D) approach that eliminates the possibility for filaments to take wrong turns at junctions by spatially separating the channels crossing each other. Specifically, 3D junctions with tunnels and overpasses were manufactured on glass substrates by two-photon polymerization, a 3D fabrication technology where a tightly focused, femtosecond-pulsed laser is scanned in a layer-to-layer fashion across a photo-polymerizable inorganic-organic hybrid polymer (ORMOCER(R)) with mu m resolution. Solidification of the polymer was confined to the focal volume, enabling the manufacturing of arbitrary 3D microstructures according to computer-aided design data. Successful realization of the 3D junction design was verified by optical and electron microscopy. Most importantly, we demonstrated the reliable transport of filaments, namely microtubules propelled by kinesin-1 motors, across these 3D junctions without junction errors. Our results open up new possibilities for 3D functional elements in biomolecular transport systems, in particular their implementation in biocomputational networks.
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| 4. |
- van Delft, Falco C. M. J. M., et al.
(författare)
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Roadmap for network-based biocomputation
- 2022
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Ingår i: Nano Futures. - : Institute of Physics Publishing (IOPP). - 2399-1984. ; 6:3
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Forskningsöversikt (refereegranskat)abstract
- Network-based biocomputation (NBC) is an alternative, parallel computation approach that can potentially solve technologically important, combinatorial problems with much lower energy consumption than electronic processors. In NBC, a combinatorial problem is encoded into a physical, nanofabricated network. The problem is solved by biological agents (such as cytoskeletal filaments driven by molecular motors) that explore all possible pathways through the network in a massively parallel and highly energy-efficient manner. Whereas there is currently a rapid development in the size and types of problems that can be solved by NBC in proof-of-principle experiments, significant challenges still need to be overcome before NBC can be scaled up to fill a technological niche and reach an industrial level of manufacturing. Here, we provide a roadmap that identifies key scientific and technological needs. Specifically, we identify technology benchmarks that need to be reached or overcome, as well as possible solutions for how to achieve this. These include methods for large-scale production of nanoscale physical networks, for dynamically changing pathways in these networks, for encoding information onto biological agents, for single-molecule readout technology, as well as the integration of each of these approaches in large-scale production. We also introduce figures of merit that help analyze the scalability of various types of NBC networks and we use these to evaluate scenarios for major technological impact of NBC. A major milestone for NBC will be to increase parallelization to a point where the technology is able to outperform the current run time of electronic processors. If this can be achieved, NBC would offer a drastic advantage in terms of orders of magnitude lower energy consumption. In addition, the fundamentally different architecture of NBC compared to conventional electronic computers may make it more advantageous to use NBC to solve certain types of problems and instances that are easy to parallelize. To achieve these objectives, the purpose of this roadmap is to identify pre-competitive research domains, enabling cooperation between industry, institutes, and universities for sharing research and development efforts and reducing development cost and time.
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