دورية أكاديمية
Development of a Screening Platform to Identify Small Molecules That Modify ELP1 Pre-mRNA Splicing in Familial Dysautonomia.
| العنوان: | Development of a Screening Platform to Identify Small Molecules That Modify ELP1 Pre-mRNA Splicing in Familial Dysautonomia. |
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| المؤلفون: | Salani M; 1 Center for Genomic Medicine, Massachusetts General Hospital Research Institute, Boston, MA, USA., Urbina F; 2 Department of Cell Biology and Physiology, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA., Brenner A; 1 Center for Genomic Medicine, Massachusetts General Hospital Research Institute, Boston, MA, USA., Morini E; 1 Center for Genomic Medicine, Massachusetts General Hospital Research Institute, Boston, MA, USA.; 3 Department of Neurology, Massachusetts General Hospital Research Institute and Harvard Medical School, Boston, MA, USA., Shetty R; 1 Center for Genomic Medicine, Massachusetts General Hospital Research Institute, Boston, MA, USA., Gallagher CS; 3 Department of Neurology, Massachusetts General Hospital Research Institute and Harvard Medical School, Boston, MA, USA., Law EA; 1 Center for Genomic Medicine, Massachusetts General Hospital Research Institute, Boston, MA, USA., Sunshine S; 1 Center for Genomic Medicine, Massachusetts General Hospital Research Institute, Boston, MA, USA., Finneran DJ; 4 Byrd Alzheimer's Institute College of Medicine Department of Molecular Pharmacology & Physiology, University of South Florida, Tampa, FL, USA., Johnson G; 5 NuPharmAdvise LLC, Sanbornton, NH, USA., Minor L; 6 In Vitro Strategies LLC, Flemington, NJ, USA., Slaugenhaupt SA; 1 Center for Genomic Medicine, Massachusetts General Hospital Research Institute, Boston, MA, USA.; 3 Department of Neurology, Massachusetts General Hospital Research Institute and Harvard Medical School, Boston, MA, USA. |
| المصدر: | SLAS discovery : advancing life sciences R & D [SLAS Discov] 2019 Jan; Vol. 24 (1), pp. 57-67. Date of Electronic Publication: 2018 Aug 07. |
| نوع المنشور: | Journal Article; Research Support, N.I.H., Extramural |
| اللغة: | English |
| بيانات الدورية: | Publisher: SAGE Publications Country of Publication: United States NLM ID: 101697563 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2472-5560 (Electronic) Linking ISSN: 24725552 NLM ISO Abbreviation: SLAS Discov Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Thousand Oaks, CA : SAGE Publications, [2017]- |
| مواضيع طبية MeSH: | Drug Evaluation, Preclinical/*methods , Dysautonomia, Familial/*genetics , RNA Precursors/*genetics , RNA Splicing/*drug effects , RNA, Messenger/*genetics , Small Molecule Libraries/*pharmacology , Transcriptional Elongation Factors/*genetics, Cell Line ; Cytokinins/pharmacology ; Exons/drug effects ; Exons/genetics ; HEK293 Cells ; Humans ; Kinetin/pharmacology ; RNA Splicing/genetics |
| مستخلص: | Familial dysautonomia (FD) is an autonomic and sensory neuropathy caused by a mutation in the splice donor site of intron 20 of the ELP1 gene. Variable skipping of exon 20 leads to a tissue-specific reduction in the level of ELP1 protein. We have shown that the plant cytokinin kinetin is able to increase cellular ELP1 protein levels in vivo and in vitro through correction of ELP1 splicing. Studies in FD patients determined that kinetin is not a practical therapy due to low potency and rapid elimination. To identify molecules with improved potency and efficacy, we developed a cell-based luciferase splicing assay by inserting renilla (Rluc) and firefly (Fluc) luciferase reporters into our previously well-characterized ELP1 minigene construct. Evaluation of the Fluc/Rluc signal ratio enables a fast and accurate way to measure exon 20 inclusion. Further, we developed a secondary assay that measures ELP1 splicing in FD patient-derived fibroblasts. Here we demonstrate the quality and reproducibility of our screening method. Development and implementation of this screening platform has allowed us to efficiently screen for new compounds that robustly and specifically enhance ELP1 pre-mRNA splicing. |
| فهرسة مساهمة: | Keywords: compounds; familial dysautonomia; luciferase assay; splicing assay |
| المشرفين على المادة: | 0 (Cytokinins) 0 (Elp1 protein, human) 0 (RNA Precursors) 0 (RNA, Messenger) 0 (Small Molecule Libraries) 0 (Transcriptional Elongation Factors) P39Y9652YJ (Kinetin) |
| تواريخ الأحداث: | Date Created: 20180808 Date Completed: 20200330 Latest Revision: 20220830 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1177/2472555218792264 |
| PMID: | 30085848 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 2472-5560 |
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| DOI: | 10.1177/2472555218792264 |