Development of a Microscale Thermophoresis assay to Target RNA Conformational Ensemble via Small Molecules against Splicing-linked Disease

Diplôme Master Development of a Microscale Thermophoresis assay to Target RNA Conformational Ensemble via Small Molecules against Splicing-linked Disease eng 2021-01-03 author Bohnet Marc Hoangphuc 876038 director Scapozza Leonardo 600318 Hutchinson–Gilford Progeria Syndrome (HGPS) is a rare and autosomal recessive genetic disorder resulting in accelerated aging in affected patients. In this condition, a single point mutation promotes the usage of an alternative splice site in exon 11 of the LMNA gene. This mutation results in a truncated protein called progerin, which is responsible for the symptoms of the disease. Previous studies have demonstrated that small molecules are able to change splicing patterns. However, there is not enough scientific report for progerin RNA and small molecule binders. In this regard, we aim to understand and to ameliorate a binding assay in order to detect small molecule binders to our target RNA hairpin structure. To reach our aim, 50 compounds from RNA binding small molecule libraries were filtered by molecular docking in silico. We also developed Microscale thermophoresis (MST) assay that shows that compounds could bind to target RNA hairpin. This method could detect possible binding interaction of filtered compounds with LMNA exon 11 cryptic splice site. The results give the first insights into the relationship between compounds and the target RNA hairpin. Further assays are required to understand splicing modulation of selected compounds binding to the target RNA hairpin. Yet this work would provide detection of small molecule binders from different libraries, besides developing a binding assay could provide detection of small molecule binders to other RNA structures in other splicing-linked diseases. Dewey 615 Master en Pharmacie 31 Section des sciences pharmaceutiques 17 false 880 Biochimie pharmaceutique 5ce7f2fb-5e5b-455f-9dd2-1b275fcac461 Master thesis Restricted 201223133022___TPR.pdf 5289544 application/pdf restrictedAccess Faculté des sciences / Section des sciences pharmaceutiques

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Development of a Microscale Thermophoresis assay to Target RNA Conformational Ensemble via Small Molecules against Splicing-linked Disease				2021	71	2

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Bohnet, Marc Hoangphuc