The advent of RNA interference as a mechanism for post-transcriptional silencing of gene function in mammalian cells represents a quantum advance and tremendous opportunity for experimental analysis of gene function. With the development of genome-wide libraries of siRNAs and shRNAs, we have entered a new era in which the application of powerful forward genetic approaches can be used both in vivo and in vitro to assign function to genes, delineate molecular pathways in which these genes affect normal and disease cellular processes, and contribute to the knowledge necessary to develop new and improve existing therapies. The Duke Center for RNA Biology, the Duke Comprehensive Cancer Center, and the Duke Institute for Genome Sciences and Policy, have come together to establish an RNAi shared-resource facility. A primary mission of the RNAi Facility is to develop and deploy RNAi technologies to support functional genomics research programs at Duke. Through large investments in RNAi and complementary technologies, we are assembling a state-of-the-art infrastructure for functional genomics, providing researchers access to genome- wide RNAi reagents and the infrastructure necessary to conduct large-scale loss-of-function studies. In this proposal, we seek support to significantly enhance this technology infrastructure with the acquisition of the Velocity 11 Integrated Instrument System for Maintenance and Delivery of RNAi/Small Molecule Libraries. This instrument system will provide the automation required for RNAi library formatting, plate handling, liquid dispensing, plate sealing and plate identification/management. This instrument will complete the suite of tools necessary to make whole genome RNAi based screening a reality for the Duke research community. The Velocity 11 system was designed from the ground up as a fully integrated computer controlled instrument and is pre-configured by Velocity 11 for the multiple workflow options necessary to maintain and deliver siRNA libraries via lipid based transfection or viral based transduction of mammalian cells while maintaining the flexibility to be useful in small molecule library screening. Coupled with our existing downstream High-content cell based imaging system, whole genome RNAi based screening represents a tremendous technological advance, enabling researchers to identify genes with roles in virtually any normal or disease-associated cellular process. ? ? ?
| Barrows, Nicholas J; Le Sommer, Caroline; Garcia-Blanco, Mariano A et al. (2010) Factors affecting reproducibility between genome-scale siRNA-based screens. J Biomol Screen 15:735-47 |
| Sessions, October M; Barrows, Nicholas J; Souza-Neto, Jayme A et al. (2009) Discovery of insect and human dengue virus host factors. Nature 458:1047-50 |
