Two-color Fluorescent Reporter

About

Background: Prior to translation, transcription generates a precursor molecule (pre-mRNA) that contains both introns (intervening sequences) and exons (protein coding regions). Alternative splicing pathways vary the production of a mature mRNA strand by modifying the introns removed and the exons joined. Depending on the splice sites, these mRNA variances give rise to proteome diversity by changing the encoded protein structure, which in turn can affect ligand binding, allosteric regulation, protein localization, etc. Although mutations in splice signals account for 15% of genetic diseases caused by point mutations indicating a pressing need for research into the mechanisms controlling alternative splicing, experimental efforts to discover compounds targeting splicing are hampered by a lack of reliable, reproducible, and high-throughput techniques.    Innovation: Researchers at UCLA have developed a reporter assay for use in the detection of compounds that modulate alternative splicing. This novel and robust two-color fluorescent system provides a means to distinguish between general changes in splicing, such as efficiency, accuracy and transcription/translation abnormalities, and a particular alternative splicing event.    Applications: Monitor alternative splicing events in individual living cells, heterogeneous cell populations and live animals.    Advantages: High-throughput  Dynamic range of applications, both in vitro and in vivo  Increased specificity to separate out alternative splicing events from other cellular processes  The inclusion of a second reporter system significantly widens detection range  Enhanced versatility due to alternative exon cassette and florescent protein swapping