Origami-SEQ · SINGLE-MOLECULE DNA SEQUENCING THROUGH DNA ORIGAMI NANOANTENNAS.
„Хоризонт 2020“ — Действия „Мария Склодовска-Кюри“
- Период
- 2017-04-01 → 2019-03-31
- Финансиране от ЕС
- 159 461 €
- Участници
- 2
- Схема
- MSCA-IF-EF-ST
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Накратко на български
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Кратко обяснение, генерирано от езиков модел по текста на CORDIS. Оригиналът е по-долу.
Резултати накратко
SINGLE-MOLECULE DNA SEQUENCING THROUGH DNA ORIGAMI NANOANTENNAS.
The objective of OrigamiSeq is to create an experimental platform that will allow scientists to study how individual biomolecules works. Among the most important techniques that allow us to study isolated biomolecules are single-molecule fluorescence techniques. One of the main challenges that faces the application of single molecule fluorescence techniques in biology is the concentration barrier. This refers to the big difference in concentration required to detect a single molecule and the concentration at which biomolecules work. The density of molecules demanded for biomolecules to work efficiently are usually at least 1000 times higher than that required to achieve single molecule detection. This means that if we use the biomolecules, or their substrate, at their working concentration, we will detect more than one molecule at a time, therefore losing the single molecule information we are seeking to obtain. To solve this problem we plan to use DNA origami nanoantennas. It is known, due to a combination of physical and chemical-physical phenomena, that a fluorescent molecule placed in the vicinity of a metal can experience an enhancement of its fluorescent emission. On the other hand, the DNA origami technique permits to build nanometer size structures with virtually any kind of shape and functionality. Our group combine both technologies to build nanostructures that can take advantage of this effect to increase the fluorescence signal coming from a single fluorescent molecule. The heart of the so-called DNA origami nanoantenna are two metal nanoparticles brought in proximity, combined with the ability to place selectively molecules in the hotspot where the enhancement is the highest. Since the signal coming from the molecule in the hotspot is far more intense that the signal coming from molecules out of it, metallic nanoantennas allow the detection of a single molecules even when several molecules at present at a time. Using this idea, we will use DNA origami nanotechnology to construct nanoantennas and selectively immobilize a biomolecule in the hotspot to study its behavior at concentrations compatible with their working concentration. We plan to develop a single-molecule DNA sequence technique in order to probe the viability of our idea. The long-term objective is to provide to the scientific community with a general platform with a high versatility that can be used to study a huge variety of biomolecules. In order to understand how they perform their work, it is of crucial importance to study the molecules that plays key roles in life at the single molecule level. Better understanding of their function will lead us to better understanding of key features of life but also on how their malfunction is related to disease. Therefore paving the way to develop better future treatments for diseases. Besides, the project includes the development of a single-molecule DNA sequencing technology, with potential applications in diagnosis.
Текст от CORDIS, на английски · Данни: CORDIS, © Европейски съюз
Цел на проекта
The inner working of many fundamental biological nanomachines implies many stochastic changes in conformation and molecular interactions. These changes are reflected as variability in the activity of the same molecule over time, and in heterogeneous activities between different molecules. By the direct observation at the molecular scale of these nanomachines activity, single-molecule fluorescence techniques have gained access to this stochastic information, which is otherwise missed by bulk techniques but is essential for their understanding. However, the widespread use of these techniques in many biological systems has been hampered by the low working concentration (nM) determined by the diffraction limit of light, and current nanophotonic solutions to this problem are technically too demanding. Here, we propose self-assembled DNA origami nanoantennas as nanophotonic platforms aiming to break this concentration barrier by means of fluorescence signal enhancing and reduction of the observation volume. The versatility of DNA origami structures, biocompatibility and ability for site-directed immobilization of biomolecules, make them perfectly suited to perform complex biological assays. In the presented action we aim to achieve single-molecule fluorescence DNA sequencing using DNA origami nanoantennas to probe the potential of these platforms to perform complex bioassays, in the high concentration regime and demanding multiplexing. Moreover, since they avoid the fabrication and instrumental challenges related to other nanophotonic devices, self-assembly DNA origami nanoantennas are amenable, easy to handle and friendly technology to the biological experimenter, and thus we expect to boost their use in biology. Besides, the fulfilment of this action will provide the fellow with a complete formative training that would boost his future scientific career, and will generate also a new DNA-sequencing technology that will impact positively European scientific excellence.
Оригинален текст от CORDIS (на английски).
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Данни: CORDIS, © Европейски съюз