Research and Development Unit, Department of Human Genetics, National Institute of Health Doutor Ricardo Jorge, INSA I.P., Rua Alexandre Herculano, 321, 4000-055 Porto, Portugal.
Center for the Study of Animal Science, Institute of Sciences, Technologies and Agro-Environment, CECA-ICETA, University of Porto, Praça Gomes Teixeira, Apartado 55142, 4051-401 Porto, Portugal.
Int J Mol Sci. 2023 Sep 27;24(19):14617. doi: 10.3390/ijms241914617.
Splicing of pre-mRNA is a crucial regulatory stage in the pathway of gene expression. The majority of human genes that encode proteins undergo alternative pre-mRNA splicing and mutations that affect splicing are more prevalent than previously thought. Targeting aberrant RNA(s) may thus provide an opportunity to correct faulty splicing and potentially treat numerous genetic disorders. To that purpose, the use of engineered U1 snRNA (either modified U1 snRNAs or exon-specific U1s-ExSpeU1s) has been applied as a potentially therapeutic strategy to correct splicing mutations, particularly those affecting the 5' splice-site (5'ss). Here we review and summarize a vast panoply of studies that used either modified U1 snRNAs or ExSpeU1s to mediate gene therapeutic correction of splicing defects underlying a considerable number of genetic diseases. We also focus on the pre-clinical validation of these therapeutic approaches both in vitro and in vivo, and summarize the main obstacles that need to be overcome to allow for their successful translation to clinic practice in the future.
前体 mRNA 的剪接是基因表达途径中的一个关键调控阶段。大多数编码蛋白质的人类基因都经历了选择性的前体 mRNA 剪接，而影响剪接的突变比以前认为的更为普遍。因此，靶向异常 RNA 可能为纠正错误剪接并治疗许多遗传疾病提供机会。为此，已经应用了工程化 U1 snRNA（修饰的 U1 snRNA 或特异性外显子 U1s-ExSpeU1s）作为一种潜在的治疗策略来纠正剪接突变，特别是那些影响 5'剪接位点（5'ss）的突变。在这里，我们回顾并总结了大量使用修饰的 U1 snRNA 或 ExSpeU1s 来介导基因治疗纠正许多遗传疾病的剪接缺陷的研究。我们还重点介绍了这些治疗方法在体外和体内的临床前验证，并总结了需要克服的主要障碍，以便将来成功将其转化为临床实践。
