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A light-responsive RNA aptamer for an azobenzene derivative.

Lotz, Thea S. ; Halbritter, Thomas ; Kaiser, Christoph ; Rudolph, Martin M. ; Kraus, Leon ; Groher, Florian ; Steinwand, Sabrina ; Wachtveitl, Josef ; Heckel, Alexander ; Suess, Beatrix (2019)
A light-responsive RNA aptamer for an azobenzene derivative.
In: Nucleic acids research, 47 (4)
doi: 10.1093/nar/gky1225
Artikel, Bibliographie

Kurzbeschreibung (Abstract)

Regulation of complex biological networks has proven to be a key bottleneck in synthetic biology. Interactions between the structurally flexible RNA and various other molecules in the form of riboswitches have shown a high-regulation specificity and efficiency and synthetic riboswitches have filled the toolbox of devices in many synthetic biology applications. Here we report the development of a novel, small molecule binding RNA aptamer, whose binding is dependent on light-induced change of conformation of its small molecule ligand. As ligand we chose an azobenzene because of its reliable photoswitchability and modified it with chloramphenicol for a better interaction with RNA. The synthesis of the ligand 'azoCm' was followed by extensive biophysical analysis regarding its stability and photoswitchability. RNA aptamers were identified after several cycles of in vitro selection and then studied regarding their binding specificity and affinity toward the ligand. We show the successful development of an RNA aptamer that selectively binds to only the trans photoisomer of azoCm with a KD of 545 nM. As the aptamer cannot bind to the irradiated ligand (λ = 365 nm), a light-selective RNA binding system is provided. Further studies may now result in the engineering of a reliable, light-responsible riboswitch.

Typ des Eintrags: Artikel
Erschienen: 2019
Autor(en): Lotz, Thea S. ; Halbritter, Thomas ; Kaiser, Christoph ; Rudolph, Martin M. ; Kraus, Leon ; Groher, Florian ; Steinwand, Sabrina ; Wachtveitl, Josef ; Heckel, Alexander ; Suess, Beatrix
Art des Eintrags: Bibliographie
Titel: A light-responsive RNA aptamer for an azobenzene derivative.
Sprache: Englisch
Publikationsjahr: 28 Februar 2019
Titel der Zeitschrift, Zeitung oder Schriftenreihe: Nucleic acids research
Jahrgang/Volume einer Zeitschrift: 47
(Heft-)Nummer: 4
DOI: 10.1093/nar/gky1225
Kurzbeschreibung (Abstract):

Regulation of complex biological networks has proven to be a key bottleneck in synthetic biology. Interactions between the structurally flexible RNA and various other molecules in the form of riboswitches have shown a high-regulation specificity and efficiency and synthetic riboswitches have filled the toolbox of devices in many synthetic biology applications. Here we report the development of a novel, small molecule binding RNA aptamer, whose binding is dependent on light-induced change of conformation of its small molecule ligand. As ligand we chose an azobenzene because of its reliable photoswitchability and modified it with chloramphenicol for a better interaction with RNA. The synthesis of the ligand 'azoCm' was followed by extensive biophysical analysis regarding its stability and photoswitchability. RNA aptamers were identified after several cycles of in vitro selection and then studied regarding their binding specificity and affinity toward the ligand. We show the successful development of an RNA aptamer that selectively binds to only the trans photoisomer of azoCm with a KD of 545 nM. As the aptamer cannot bind to the irradiated ligand (λ = 365 nm), a light-selective RNA binding system is provided. Further studies may now result in the engineering of a reliable, light-responsible riboswitch.

ID-Nummer: pmid:30517682
Fachbereich(e)/-gebiet(e): 10 Fachbereich Biologie
10 Fachbereich Biologie > Synthetic Genetic Circuits (2020 umbenannt in "Synthetic RNA biology")
Hinterlegungsdatum: 10 Dez 2018 11:47
Letzte Änderung: 06 Jun 2019 10:28
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