Department of Genetics, Harvard Medical School, Boston, Massachusetts 02115.
Genetics. 2013 Sep;195(1):37-45. doi: 10.1534/genetics.113.151340. Epub 2013 Jun 21.
RNA interference (RNAi) is a widely adopted tool for loss-of-function studies but RNAi results only have biological relevance if the reagents are appropriately mapped to genes. Several groups have designed and generated RNAi reagent libraries for studies in cells or in vivo for Drosophila and other species. At first glance, matching RNAi reagents to genes appears to be a simple problem, as each reagent is typically designed to target a single gene. In practice, however, the reagent-gene relationship is complex. Although the sequences of oligonucleotides used to generate most types of RNAi reagents are static, the reference genome and gene annotations are regularly updated. Thus, at the time a researcher chooses an RNAi reagent or analyzes RNAi data, the most current interpretation of the RNAi reagent-gene relationship, as well as related information regarding specificity (e.g., predicted off-target effects), can be different from the original interpretation. Here, we describe a set of strategies and an accompanying online tool, UP-TORR (for Updated Targets of RNAi Reagents; www.flyrnai.org/up-torr), useful for accurate and up-to-date annotation of cell-based and in vivo RNAi reagents. Importantly, UP-TORR automatically synchronizes with gene annotations daily, retrieving the most current information available, and for Drosophila, also synchronizes with the major reagent collections. Thus, UP-TORR allows users to choose the most appropriate RNAi reagents at the onset of a study, as well as to perform the most appropriate analyses of results of RNAi-based studies.
RNA 干扰 (RNAi) 是一种广泛应用于功能丧失研究的工具,但只有当试剂被适当地映射到基因上时,RNAi 结果才具有生物学相关性。几个小组已经设计并生成了用于在细胞内或体内进行果蝇和其他物种研究的 RNAi 试剂库。乍一看,将 RNAi 试剂与基因匹配似乎是一个简单的问题,因为每个试剂通常都被设计靶向单个基因。然而,在实践中,试剂与基因的关系是复杂的。尽管用于生成大多数类型的 RNAi 试剂的寡核苷酸序列是静态的,但参考基因组和基因注释经常会更新。因此,当研究人员选择 RNAi 试剂或分析 RNAi 数据时,当前对 RNAi 试剂-基因关系的最准确解释,以及有关特异性的相关信息(例如,预测的脱靶效应),可能与最初的解释不同。在这里,我们描述了一组策略和一个配套的在线工具,UP-TORR(用于更新 RNAi 试剂的靶标;www.flyrnai.org/up-torr),用于准确和最新的基于细胞和体内 RNAi 试剂的注释。重要的是,UP-TORR 每天自动与基因注释同步,检索最新的可用信息,对于果蝇,也与主要试剂库同步。因此,UP-TORR 允许用户在研究开始时选择最合适的 RNAi 试剂,以及对基于 RNAi 的研究结果进行最合适的分析。