title: High-throughput analysis of an RNAi library identifies novel kinase targets in Trypanosoma brucei.
abstract: New drugs are needed to treat human African trypanosomiasis because the currently approved treatments are toxic or limited in efficacy. One strategy for developing new drugs involves discovering novel genes whose products can be targeted for modulation by small-molecule chemotherapeutic agents. The Trypanosoma brucei genome contains many genes with the potential to become such targets. Kinases represent one group of genes that regulate many important cell functions and can be modulated by small molecules, thus representing a promising group of enzymes to screen for potential therapeutic targets. RNAi screens could help identify the most promising kinase targets, but the lack of suitable assays represents a barrier for optimizing the use of this technology in T. brucei. Here, we describe an RNAi screen of a small RNAi library targeting 30 members of the T. brucei kinome utilizing a luciferase-based assay. This screen both validated the luciferase-based assay as a suitable method for conducting RNAi screens in T. brucei and also identified two kinases (CRK12 and ERK8) that are essential for normal proliferation by the parasite.
authors.affiliation: Department of Pathology, Sandler Center for Drug Discovery, University of California, San Francisco, CA 94158-2330, USA.
authors.forename: Zachary B
authors.initials: ZB
authors.lastname: Mackey
authors.forename: Kyriacos
authors.initials: K
authors.lastname: Koupparis
authors.forename: Mari
authors.initials: M
authors.lastname: Nishino
authors.forename: James H
authors.initials: JH
authors.lastname: McKerrow
mesh_terms: D000818:Animals; D055808:Drug Discovery; D006801:Humans; D010770:Phosphotransferases / Q000235:genetics*; D034622:RNA Interference; D014346:Trypanosoma brucei brucei / Q000201:enzymology* / Q000235:genetics; D014353:Trypanosomiasis, African / Q000188:drug therapy
chemical_list: D010770:Phosphotransferases