Broccoli Fluorets: Split Aptamers as a User-Friendly Fluorescent Toolkit for Dynamic RNA Nanotechnology

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dc.contributor.authorChandler, Morganen
dc.contributor.authorLyalina, Tatianaen
dc.contributor.authorHalman, Justinen
dc.contributor.authorRackley, Laurenen
dc.contributor.authorLee, Laurenen
dc.contributor.authorDang, Dylanen
dc.contributor.authorKe, Weinaen
dc.contributor.authorSajja, Sameeren
dc.contributor.authorWoods, Stevenen
dc.contributor.authorAcharya, Shrijaen
dc.contributor.authorBaumgarten, Elijahen
dc.contributor.authorChristopher, Jonathanen
dc.contributor.authorElshalia, Emmanen
dc.contributor.authorHrebien, Gabrielen
dc.contributor.authorKublank, Kinzeyen
dc.contributor.authorSaleh, Sajaen
dc.contributor.authorStallings, Baileyen
dc.contributor.authorTafere, Michaelen
dc.contributor.authorStriplin, Carynen
dc.contributor.authorAfonin, Kirill A.en
dc.contributor.departmentBiochemistryen
dc.date.accessioned2018-12-21T15:38:46Zen
dc.date.available2018-12-21T15:38:46Zen
dc.date.issued2018-12-02en
dc.date.updated2018-12-21T14:54:20Zen
dc.description.abstractRNA aptamers selected to bind fluorophores and activate their fluorescence offer a simple and modular way to visualize native RNAs in cells. Split aptamers which are inactive until the halves are brought within close proximity can become useful for visualizing the dynamic actions of RNA assemblies and their interactions in real time with low background noise and eliminated necessity for covalently attached dyes. Here, we design and test several sets of F30 Broccoli aptamer splits, that we call fluorets, to compare their relative fluorescence and physicochemical stabilities. We show that the splits can be simply assembled either through one-pot thermal annealing or co-transcriptionally, thus allowing for direct tracking of transcription reactions via the fluorescent response. We suggest a set of rules that enable for the construction of responsive biomaterials that readily change their fluorescent behavior when various stimuli such as the presence of divalent ions, exposure to various nucleases, or changes in temperature are applied. We also show that the strand displacement approach can be used to program the controllable fluorescent responses in isothermal conditions. Overall, this work lays a foundation for the future development of dynamic systems for molecular computing which can be used to monitor real-time processes in cells and construct biocompatible logic gates.en
dc.description.versionPublished versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationChandler, M.; Lyalina, T.; Halman, J.; Rackley, L.; Lee, L.; Dang, D.; Ke, W.; Sajja, S.; Woods, S.; Acharya, S.; Baumgarten, E.; Christopher, J.; Elshalia, E.; Hrebien, G.; Kublank, K.; Saleh, S.; Stallings, B.; Tafere, M.; Striplin, C.; Afonin, K.A. Broccoli Fluorets: Split Aptamers as a User-Friendly Fluorescent Toolkit for Dynamic RNA Nanotechnology. Molecules 2018, 23, 3178.en
dc.identifier.doihttps://doi.org/10.3390/molecules23123178en
dc.identifier.urihttp://hdl.handle.net/10919/86503en
dc.language.isoenen
dc.publisherMDPIen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.subjectRNA nanotechnologyen
dc.subjectaptameren
dc.subjectBroccolien
dc.subjectdynamic nanoparticlesen
dc.subjectconditional activationen
dc.titleBroccoli Fluorets: Split Aptamers as a User-Friendly Fluorescent Toolkit for Dynamic RNA Nanotechnologyen
dc.title.serialMoleculesen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

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