Stable suppression of tumorigenicity by virus-mediated RNA interference. The ink4a/ARF network in tumour suppression. Duplexes of 21-nucleotide RNAs mediate RNA interference in cultured mammalian cells. Short hairpin RNAs (shRNAs) induce sequence-specific silencing in mammalian cells. A system for stable expression of short interfering RNAs in mammalian cells. Genome-wide RNAi analysis of Caenorhabditis elegans fat regulatory genes. Systematic functional analysis of the Caenorhabditis elegans genome using RNAi. Identification of Hedgehog pathway components by RNAi in Drosophila cultured cells. These new tools will greatly facilitate large-scale loss-of-function genetic screens in mammalian cells. Furthermore, we describe siRNA bar-code screens to rapidly identify individual siRNA vectors associated with a specific phenotype. Suppression of these genes confers resistance to both p53-dependent and p19 ARF-dependent proliferation arrest, and abolishes a DNA-damage-induced G1 cell-cycle arrest. We use this RNAi library in human cells to identify one known and five new modulators of p53-dependent proliferation arrest. Here we report the construction of a set of retroviral vectors encoding 23,742 distinct shRNAs, which target 7,914 different human genes for suppression. We and others have recently developed expression vectors to direct the synthesis of short hairpin RNAs (shRNAs) that act as short interfering RNA (siRNA)-like molecules to stably suppress gene expression 4, 5.

In mammalian cells, such screens have been hampered by a lack of suitable tools that can be used on a large scale.

RNA interference (RNAi) is a powerful new tool with which to perform loss-of-function genetic screens in lower organisms and can greatly facilitate the identification of components of cellular signalling pathways 1, 2, 3.

Pathway download the new