Antisense Methods to Modulate Pre-mRNA Splicing
The dynamic process of pre-mRNA splicing is regulated by combinatorial control exerted by overlapping cis-elements that are unique to every exon and its flanking intronic sequences. Splicing cis-elements are usually 4–8-nucleotide-long linear motifs that furnish interaction sites for specific proteins. Secondary and higher-order RNA structures exert an additional layer of control by providing accessibility to cis-elements. Antisense oligonucleotides (ASOs) that block splicing cis-elements and/or affect RNA structure have been shown to modulate alternative splicing in vivo. Consistently, ASO-based strategies have emerged as a powerful tool for therapeutic manipulation of aberrant splicing in pathological conditions. Here we describe the application of an ASO-based approach for the enhanced production of the full-length mRNA of SMN2 in spinal muscular atrophy patient cells.
Key wordsAntisense oligonucleotide (ASO) Survival motor neuron (SMN) Pre-mRNA splicing Multi-exon-skipping detection assay (MESDA) Intronic splicing silencer N1 (ISS-N1) GC-rich sequence GM03813 Spinal muscular atrophy (SMA) Phosphorothioate 2′-O-methyl modification Transfection
This work was supported by grants from United States National Institutes of Health (R01 NS055925, R21 NS072259, and R21 NS080294) and Salsbury Endowment (Iowa State University, Ames, IA, USA) to RNS. Authors acknowledge Dr. Natalia Singh for providing critical comments on the manuscript.