Mise au point d'une méthode de mesure d'interaction ligand-ARN par électrochimie

Abstract : RNA molecules play a major role in various biochemical processes and they are now considered as an important drug target. However, our limited understanding of the interactions occurring between small molecules and RNA complicate the search for new ligands (or drugs) with improved specific interaction and binding to elaborated RNA structures. In the absence of any rational approach, a screening strategy could shed light on the ligand/RNA interactions. In this thesis, we describe a simple electrochemical approach allowing for high-throughput detection and quantification of small molecule/RNA interactions. The principle of the method relies on the difference of diffusion rates between a redoxmolecular probe free or bound to its RNA target and thus to the ability to more easily electrochemically detect the forme rover the latter in a homogenous solution. This electrochemical detection strategy has the advantages of being affordable,fast, easy to use, sensitive and well-adapted to a high-throughput screening strategy in small volume samples. This methodology was used to characterize the binding of an aminoglycoside analog bearing a ferocenyl group to the ribosomal RNA fragment (rRNA 16S23). Furthermore, competitive binding of unlabelled aminoglycosides on theRNA/electrochemical probe complex allowed us to evaluate their dissociation constants (KD). These competitive experiments could further be generalized to measure KD values for libraries of molecules, which could help to find better RNA ligands.
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Hélène Guyon. Mise au point d'une méthode de mesure d'interaction ligand-ARN par électrochimie. Biochimie [q-bio.BM]. Université Sorbonne Paris Cité, 2016. Français. ⟨NNT : 2016USPCB147⟩. ⟨tel-01978016⟩

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