REAGENTS AND METHODS FOR IDENTIFICATION OF RNAi PATHWAY GENES AND CHEMICAL MODULATORS OF RNAi

摘要

The present invention provides reagents such as cells, cell lines, and vectors, that can be used to identify mammalian genes whose expression products (RNA or protein) play a role in RNA interference (RNAi) and/or to identify chemical modulators of RNAi, or for other purposes. The invention further provides a variety of methods for identifying such genes or modulators. In particular, the invention provides a mammalian cell comprising a nucleic acid that encodes a selectable marker and one or more nucleic acid templates for transcription of an RNAi-inducing agent integrated into the genome of the cell, wherein the RNAi-inducing agent reduces expression of the marker and is not naturally found in the cell. Additional cells and cell lines comprising nucleic acids that encode one or more additional markers are also provided. According to certain of the inventive methods cells such as these are mutagenized, transfected or infected with a library of genetic suppressor elements, or contacted with a test compound. Cells in which RNAi is inhibited or activated are identified using an appropriate selective condition or screening method. The identity of the mutated or inhibited gene or the identity of the compound is then determined.