Dery Kenneth J, Gusti Veronica, Gaur Shikha, Shively John E, Yen Yun, Gaur Rajesh K
Division of Molecular Biology, Beckman Research Institute of the City of Hope, Duarte, CA, USA.
Methods Mol Biol. 2009;555:127-44. doi: 10.1007/978-1-60327-295-7_10.
The majority of eukaryotic genes undergo alternative splicing, an evolutionarily conserved phenomenon, to generate functionally diverse protein isoforms from a single transcript. The fact that defective pre-mRNA splicing can generate non-functional and often toxic proteins with catastrophic effects, accurate removal of introns and joining of exons is vital for cell homeostasis. Thus, molecular tools that could either silence a disease-causing gene or regulate its expression in trans will find many therapeutic applications. Here we present two RNA-based approaches, namely RNAi and theophylline-responsive riboswitch that can regulate gene expression by loss-of-function and modulation of splicing, respectively. These strategies are likely to continue to play an integral role in studying gene function and drug discovery.
大多数真核基因会经历可变剪接，这是一种进化上保守的现象，能从单个转录本产生功能多样的蛋白质异构体。由于有缺陷的前体mRNA剪接会产生无功能且通常有毒的蛋白质，从而带来灾难性影响，因此准确去除内含子和连接外显子对细胞稳态至关重要。因此，能够沉默致病基因或反式调节其表达的分子工具将有许多治疗应用。在此，我们展示了两种基于RNA的方法，即RNA干扰和茶碱响应性核糖开关，它们分别可以通过功能丧失和剪接调控来调节基因表达。这些策略可能会继续在基因功能研究和药物发现中发挥不可或缺的作用。
