A high-throughput screening assay for the functional delivery of splice-switching oligonucleotides in human melanoma...

Since the conception of RNA nanotechnology (Cell, 94:147, 1998), there has been tremendous interest in its application for the functional delivery of RNA into cells. Splice-switching oligonucleotides (SSOs) are an emerging antisense drug class with the ability to therapeutically modify gene expression. A wide variety of chemical modifications have been devised to try to increase the activity and stability of SSOs. Also, as with most nucleic acid therapeutics, delivery into the cell is the major hurdle for in vivo and clinical applications. As a result, various RNA nanoparticles are being constructed for targeted delivery of therapeutics. However, it is difficult to find a practical assay to measure splice-switching activity. Here, we describe a model delivery system that can be used as a convenient, high-throughput assay to quantitatively measure the functional delivery and splicing redirection in a live human melanoma cell line.