Multiplex in situ hybridization within a single transcript: RNAscope reveals dystrophin mRNA dynamics.

ABSTRACT: Dystrophin plays a vital role in maintaining muscle health, yet low mRNA expression, lengthy transcription time and the limitations of traditional in-situ hybridization (ISH) methodologies mean that the dynamics of dystrophin transcription remain poorly understood. RNAscope is highly sensitive ISH method that can be multiplexed, allowing detection of individual transcript molecules at sub-cellular resolution, with different target mRNAs assigned to distinct fluorophores. We instead multiplex within a single transcript, using probes targeted to the 5' and 3' regions of muscle dystrophin mRNA. Our approach shows this method can reveal transcriptional dynamics in health and disease, resolving both nascent myonuclear transcripts and exported mature mRNAs in quantitative fashion (with the latte

SUBMITTER: Hildyard JCW 

PROVIDER: S-EPMC7514052 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature