Popis: |
Exon-specific isoform expression from alternatively spliced mRNA is a fundamental mechanism that substantially enriches a cell’s proteome. However, conventional methods to assess alternative splicing are either consumptive and work-intensive or do not quantify isoform expression over multiple time points and at the protein level. We thus developed an exon-specific isoform expression reporter system (EXSISERS), which non-invasively reports the translation of exon-containing isoforms by scarlessly excising reporter proteins from the nascent polypeptide chain via highly efficient, intein-mediated protein splicing. EXSISERS enables non-invasive monitoring of exon-specific isoform expression with high sensitivity and cellular resolution and enables high-throughput screening of exon-specific therapeutic interventions.This protocol on how to generate EXSISERS reporter cell lines is related to the publication ‘Non-invasive and high-throughput interrogation of exon-specific isoform expression’ in Nature Cell Biology. |