RNA interference (RNAi) Platform, Broad Institute of Harvard and Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Nat Methods. 2011 Jun 26;8(8):659-61. doi: 10.1038/nmeth.1638.
Functional characterization of the human genome requires tools for systematically modulating gene expression in both loss-of-function and gain-of-function experiments. We describe the production of a sequence-confirmed, clonal collection of over 16,100 human open-reading frames (ORFs) encoded in a versatile Gateway vector system. Using this ORFeome resource, we created a genome-scale expression collection in a lentiviral vector, thereby enabling both targeted experiments and high-throughput screens in diverse cell types.
功能基因组学的研究要求能够系统地在基因功能丧失和获得实验中调节基因表达的工具。我们描述了一个经过序列验证的、克隆的超过 16100 个人类开放阅读框(ORF)的集合的产生,这些 ORF 编码在一个多功能的 Gateway 载体系统中。利用这个 ORFeome 资源,我们在慢病毒载体中创建了一个基因组规模的表达谱集合,从而能够在多种细胞类型中进行靶向实验和高通量筛选。