Fuchs Florian, Boutros Michael
Boveri-Group Signaling and Functional Genomics, German Cancer Research Center, Im Neuenheimer Feld 580, D-69120 Heidelberg, Germany.
Brief Funct Genomic Proteomic. 2006 Mar;5(1):52-6. doi: 10.1093/bfgp/ell007. Epub 2006 Feb 23.
A systematic characterization of genes with unknown function is a key challenge after the sequencing of the human genome and the genomes of many model organisms. High-throughput RNA-interference (RNAi) screenings have become a widely used approach in invertebrate model organisms and also promise to revolutionize cell biology in mammals. Genome-wide RNAi screens in Caenorhabditis elegans and Drosophila, and in a smaller scale in mammalian cells have proven to be a valuable and successful method for the dissection of diverse biological processes. A number of RNAi libraries have become available that rely on different technologies, such as long double-stranded (ds) RNAs, in vitro diced short-interfering (si) RNAs, synthetic siRNAs and short-hairpin (sh) RNAs, which all have specific advantages and disadvantages. In addition, progress in screening technologies and data analysis allows the adaptation of screening methods to analyse more complex cellular processes. This review will summarize strategies in combining genome-scale RNAi libraries, high-throughput screening technologies, integrated high-content data analysis and will discuss future challenges.
在人类基因组以及许多模式生物基因组测序完成之后，对功能未知基因进行系统表征是一项关键挑战。高通量RNA干扰（RNAi）筛选已成为无脊椎动物模式生物中广泛使用的方法，也有望给哺乳动物细胞生物学带来变革。在秀丽隐杆线虫和果蝇中进行的全基因组RNAi筛选，以及在哺乳动物细胞中较小规模的筛选，已被证明是剖析各种生物学过程的一种有价值且成功的方法。现在已有多种依赖不同技术的RNAi文库可供使用，比如长双链（ds）RNA、体外切割的短干扰（si）RNA、合成siRNA以及短发夹（sh）RNA，它们都各有优缺点。此外，筛选技术和数据分析方面的进展使得筛选方法能够得以调整，用于分析更复杂的细胞过程。本综述将总结在结合基因组规模RNAi文库、高通量筛选技术、综合高内涵数据分析方面的策略，并将讨论未来面临的挑战。
