1 Global Research Laboratory for RNAi Medicine, Department of Chemistry, Sungkyunkwan University , Suwon, Korea.
Nucleic Acid Ther. 2014 Jun;24(3):192-8. doi: 10.1089/nat.2013.0466. Epub 2014 Feb 14.
The gene-silencing activity of a small interfering RNA (siRNA) is determined by various factors. Considering that RNA interference (RNAi) is an unparalleled technology in both basic research and therapeutic applications, thorough understanding of the factors determining RNAi activity is critical. This report presents observations that siRNAs targeting KRT7 show cell-line-dependent activity, which correlates with the expression level of KRT7 mRNA. By modulating the target mRNA level, it was confirmed that highly expressed genes are more susceptible to siRNA-mediated gene silencing. Finally, several genes that show different expression levels in a cell-line dependent manner were tested, which verified the expression-level-dependent siRNA activities. These results strongly suggest that the abundance of target mRNA is a critical factor that determines the efficiency of the siRNA-mediated gene silencing in a given cellular context. This report should provide practical guidelines for designing RNAi experiments and for selecting targetable genes in RNAi therapeutics studies.
小干扰 RNA（siRNA）的基因沉默活性由多种因素决定。鉴于 RNA 干扰（RNAi）在基础研究和治疗应用中是一种无与伦比的技术，因此彻底了解决定 RNAi 活性的因素至关重要。本报告介绍了针对 KRT7 的 siRNA 表现出细胞系依赖性活性的观察结果，这与 KRT7 mRNA 的表达水平相关。通过调节靶 mRNA 水平，证实了高表达基因更容易受到 siRNA 介导的基因沉默的影响。最后，测试了在细胞系依赖性方式下表达水平不同的几个基因，验证了 siRNA 活性的表达水平依赖性。这些结果强烈表明靶 mRNA 的丰度是决定给定细胞环境中 siRNA 介导的基因沉默效率的关键因素。本报告应为设计 RNAi 实验以及在 RNAi 治疗学研究中选择靶向基因提供实用指南。
