RNAi Screening or Selection Studies
The availability of RNAi libraries targeting entire genomes (or large subsets) provides the opportunity to conduct loss-of-function genetic screens or selections in mammalian cells. Conceptually, there are two primary approaches to cell-based, large-scale identification of functionally relevant genes - pooled clone selection assays and highly parallel, individual clone screening assays. Each approach offers distinct advantages and limitations, and this facility will work with researchers to develop, evaluate, and implement cell-based screening and selection studies.
The RNAi Facility is developing the infrastructure to enable large-scale RNAi studies, including cell-based assay automation and analysis. We are interested in discussing your projects and learning how we might be able to contribute.
Resources available:
- Human retroviral shRNA collection: Individual clones are available for functional validation and discovery studies (for example, candidate genes, pathways, or gene classes), enabling systematic analysis of 10s to 1000s of clones in parallel or pooled assays.
- Pooled shRNA plasmid DNAs: High-quality shRNA plasmid DNAs have been prepared by independently culturing all ~62,500 clones in the Hannon-Elledge human retroviral shRNA library, pooling these cultures (sets of ~2000), and preparing transfection-grade (endotoxin-free) plasmid DNA. This DNA is available for pooled selection studies as 32 unique pools of ~2000 clones, and can be further combined as appropriate.
For more information on RNAi screening or selection studies, contact James Pearson.