DI-UMONS : Dépôt institutionnel de l’université de Mons

Recherche transversale
(titres de publication, de périodique et noms de colloque inclus)
2018-06-25 - Colloque/Présentation - communication orale - Anglais - page(s)

Neaga Ioan-Ovidiu, Hambye Stéphanie , Baroni Alexandra, Bodoki Ede, Blankert Bertrand , Oprean Radu, "Screening drug candidates for myotonic dystrophy type 1: nucleic acid probe design and method development" in 18th CEEPUS Symposium and Summer School on Bioanalysis 2018, 25 – 30 June, Komarno, Slovakia, 2018

  • Codes CREF : Chimie analytique (DI1314), Sciences pharmaceutiques (DI3400), Techniques séparatives (DI2729)
  • Unités de recherche UMONS : Analyse pharmaceutique (M130)
  • Instituts UMONS : Institut des Sciences et Technologies de la Santé (Santé)

Abstract(s) :

(Anglais) Myotonic dystrophy type 1 is an autosomal dominantly inherited degenerative disease characterized by progressive loss of muscle mass, cataract, hypersomnia, fatigue, heart conduction abnormalities and respiratory failure [1]. The mechanism of the disease can be traced back to a mutation on the site of the DMPK (dystrophia myotonica protein kinase) gene. The CTG triplet expansion present at this site is transcribed into (CUG)n RNA repeats, which in turn is able to bind splicing factors from which MBLN1 and CUGBP1. The deficiency in these factors will result in RNA mis-splicing and protein synthesis defects. While at the present there is no available treatment for the disease, several groups focused their attention on finding small molecules that could disrupt the MBNL-1/CUG complex or bind to DNA CTG repeat sequence thus preventing its transcription [2,3]. In this study we present an affinity capillary electrophoresis method along with a workflow for producing nucleic acid probes required for screening small molecules with potential activity in myotonic dystrophy type 1. For the method development and early testing of ligands, an RNA probe with 50 CUG repeats was used. In the second part of the study we focused on producing and purifying DNA and RNA fragments with lengths corresponding with those present in both physiological (14 CUG/CTG) and pathological states (95 CUG/CTG). The DNA probes were produced by bacterial amplification in E. coli. The obtained plasmid containing the desired fragment was double digested and purified using a custom-built gel electrophoresis chamber and extracted by electrodialysis. The RNA probes were produced by in vitro transcription. The obtained fragments were tested against a small library of ligands containing various antibiotics and small molecules. The developed ACE method proved to be efficient and robust, allowing the testing of a ligand in a few hours, with minimum costs of reagents.