Quellos High Throughput Screening Core
RNA Interference Screening
The combination of RNA interference (RNAi) and high throughput screening (HTS) has opened a new frontier toward the unbiased discovery of genes essential for specific biological processes or disease states. Utilizing synthetic small RNA duplexes (siRNA) or small RNA hairpins (shRNA), the latter expressed from viral vectors, specific genes are inactivated within cells. This “loss of function” assay aids in pinpointing the role specific gene products play in cellular processes. Laboratory automation renders RNAi screening easily applicable to most cell types allowing for the rapid interrogation of gene function of the entire genomes of human, mouse or other model organisms. An added potential of RNAi screening is the identification of new drug targets for any disease for which a cell based model is available (e.g., cancer, neurological or cardiac diseases). Current offerings at the Quellos High Throughput Screening Core include sub-genomic siRNA pilot screens in libraries generated from human or mouse and anticipated full genome scale siRNA/shRNA screens.
Pilot screens:
siRNAs targeting the human and mouse kinases (~800 genes) from SIGMA and Ambion. The libraries contain 3 siRNAs per target and they are pooled in a single well. Screens will be preferentially performed in 384 well format in triplicates. We also have capacity for 1,536 in case of limited biological material.
Full scale screens:
siRNAs targeting the extended druggable genome (~10,000 genes including transcription factors) for both human and mouse. Format as above.
Lentiviral sort hairpin (sh) arrayed libraries
Lentiviral sh libraries targeting the human kinases. Pools of three different sh lentiviruses are arrayed in 384 well format ready to screen (Lentiexpress, SIGMA).
Contact
Carla Grandori, M.D., Ph.D.
Director
Phone: 206-543-8119
E-mail: grandc@uw.edu
