High-throughput sensitive screening of small molecule modulators of microexon alternative splicing using dual Nano and Firefly luciferase reporters.
Nat Commun
; 15(1): 6328, 2024 Jul 27.
Article
em En
| MEDLINE
| ID: mdl-39068192
ABSTRACT
Disruption of alternative splicing frequently causes or contributes to human diseases and disorders. Consequently, there is a need for efficient and sensitive reporter assays capable of screening chemical libraries for compounds with efficacy in modulating important splicing events. Here, we describe a screening workflow employing dual Nano and Firefly luciferase alternative splicing reporters that affords efficient, sensitive, and linear detection of small molecule responses. Applying this system to a screen of ~95,000 small molecules identified compounds that stimulate or repress the splicing of neuronal microexons, a class of alternative exons often disrupted in autism and activated in neuroendocrine cancers. One of these compounds rescues the splicing of several analyzed microexons in the cerebral cortex of an autism mouse model haploinsufficient for Srrm4, a major activator of brain microexons. We thus describe a broadly applicable high-throughput screening system for identifying candidate splicing therapeutics, and a resource of small molecule modulators of microexons with potential for further development in correcting aberrant splicing patterns linked to human disorders and disease.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Éxons
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Processamento Alternativo
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Genes Reporter
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Luciferases de Vaga-Lume
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Bibliotecas de Moléculas Pequenas
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Ensaios de Triagem em Larga Escala
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article