Dallas Anne, Johnston Brian H
SomaGenics, Inc., Santa Cruz, CA, USA.
Methods Mol Biol. 2013;942:279-90. doi: 10.1007/978-1-62703-119-6_15.
Synthetic shRNAs that are too short to be Dicer substrates (short shRNAs or sshRNAs) can be highly potent RNAi effectors when properly designed, with activities similar to or more potent than the more commonly used siRNAs targeting the same sequences. sshRNAs can be designed in two possible orientations: left- or right-hand loop, designated L-sshRNAs and R-sshRNAs, respectively. Because L- and R-sshRNAs are processed by the RNAi machinery in different ways, optimal designs for the two formats diverge in several key aspects. Here, we describe the principles of design and chemical modification of highly effective L- and R-sshRNAs.
合成的短发夹RNA（shRNA）若短到无法成为Dicer酶的底物（短shRNA或sshRNA），经过合理设计后可成为高效的RNA干扰效应物，其活性与靶向相同序列的更常用的小干扰RNA（siRNA）相似或更强。sshRNA可以设计成两种可能的方向：左手环或右手环，分别称为L-sshRNA和R-sshRNA。由于L-sshRNA和R-sshRNA由RNA干扰机制以不同方式加工，这两种形式的最佳设计在几个关键方面存在差异。在此，我们描述高效L-sshRNA和R-sshRNA的设计原则及化学修饰方法。
