Department of Human Genetics and Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands.
Departments of Pediatrics and Human Genetics, Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525GA Nijmegen, The Netherlands.
Int J Mol Sci. 2021 Apr 28;22(9):4621. doi: 10.3390/ijms22094621.
The discovery of novel intronic variants in the locus has contributed significantly to solving the missing heritability in Stargardt disease (STGD1). The increasing number of variants affecting pre-mRNA splicing makes a suitable candidate for antisense oligonucleotide (AON)-based splicing modulation therapies. In this study, AON-based splicing modulation was assessed for 15 recently described intronic variants (three near-exon and 12 deep-intronic variants). In total, 26 AONs were designed and tested in vitro using a midigene-based splice system. Overall, partial or complete splicing correction was observed for two variants causing exon elongation and all variants causing pseudoexon inclusion. Together, our results confirm the high potential of AONs for the development of future RNA therapies to correct splicing defects causing STGD1.
在 基因中发现的新型内含子变异显著有助于解决斯塔加特病（STGD1）的遗传缺失问题。越来越多的影响前体 mRNA 剪接的变异使 成为反义寡核苷酸（AON）为基础的剪接调节治疗的合适候选者。在这项研究中，对 15 个最近描述的内含子变异（三个近外显子和 12 个深内含子变异）进行了基于 AON 的剪接调节评估。总共设计并测试了 26 个 AON 使用基于 midigene 的剪接系统进行体外。总体而言，对于导致外显子延长的两个变异体观察到部分或完全剪接纠正，并且所有导致假外显子包含的变异体均观察到。总之，我们的结果证实了 AON 用于开发未来 RNA 治疗以纠正导致 STGD1 的剪接缺陷的高潜力。
