HiFENS: high-throughput FISH detection of endogenous pre-mRNA splicing isoforms.

ABSTRACT: Splicing factors play an essential role in regulation of alternative pre-mRNA splicing. While much progress has been made in delineating the mechanisms of the splicing machinery, the identity of signal transduction pathways and upstream factors that regulate splicing factor activity is largely unknown. A major challenge in the discovery of upstream regulatory factors of pre-mRNA splicing is the scarcity of functional genomics screening methods to monitor splicing outcomes of endogenous genes. Here, we have developed HiFENS (high throughput FISH detection of endogenous splicing isoforms), a high-throughput imaging assay based on hybridization chain reaction (HCR) and used HiFENS to screen for cellular factors that regulate alternative splicing of endogenous genes. We demonstrate optimized d

SUBMITTER: Shilo A 

PROVIDER: S-EPMC9825148 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature