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