This protocol describes a minigene reporter assay to monitor the impact of 5´-splice site mutations on splicing and develops suppressor U1 snRNA for the rescue of mutation-induced splicing inhibition. The reporter and suppressor U1 snRNA constructs are expressed in HeLa cells, and splicing is analyzed by primer extension or RT-PCR.
Wong, J., Martelly, W., Sharma, S. A Reporter Based Cellular Assay for Monitoring Splicing Efficiency. J. Vis. Exp. (175), e63014, doi:10.3791/63014 (2021).