Singh Natalia N, Luo Diou, Singh Ravindra N
Department of Biomedical Sciences, College of Veterinary Medicine, Iowa State University, Ames, IA, USA.
Methods Mol Biol. 2025;2964:381-404. doi: 10.1007/978-1-0716-4730-1_25.
Pre-mRNA splicing, a dynamic process of intron removal and exon joining, is governed by a combinatorial control exerted by overlapping cis-elements that are unique to each exon and its flanking intronic sequences. Splicing cis-elements are usually 4- to 8-nucleotide-long linear motifs that provide binding sites for specific proteins. Pre-mRNA splicing is also influenced by secondary- and higher-order RNA structures that affect accessibility of splicing cis-elements. Antisense oligonucleotides (ASOs) that block splicing cis-elements and/or affect RNA structure have been shown to modulate splicing in vivo. Therefore, ASO-based strategies have emerged as a powerful tool for therapeutic manipulation of splicing in pathological conditions. Here we describe an ASO-based approach to increase the production of the full-length SMN2 mRNA in Spinal Muscular Atrophy patient cells.
前体mRNA剪接是一个去除内含子和连接外显子的动态过程,由重叠的顺式元件施加的组合控制所支配,这些顺式元件对于每个外显子及其侧翼内含子序列而言都是独特的。剪接顺式元件通常是4至8个核苷酸长的线性基序,为特定蛋白质提供结合位点。前体mRNA剪接还受影响剪接顺式元件可及性的二级和高级RNA结构的影响。已证明阻断剪接顺式元件和/或影响RNA结构的反义寡核苷酸(ASO)可在体内调节剪接。因此,基于ASO的策略已成为在病理条件下对剪接进行治疗性操控的强大工具。在此,我们描述了一种基于ASO的方法,以增加脊髓性肌萎缩症患者细胞中全长SMN2 mRNA的产生。