The Functional Genomics Shared Resource was established as a shared resource for the application of functional genomic technologies to support the research programs of investigators at Duke. Through investments in genetic and chemical perturbation technologies, we have assembled state-of-the-art reagents and equipment, providing researchers access to genome-wide RNAi reagents and the infrastructure necessary to conduct large-scale functional studies in mammalian cells.
- Expertise: The facility serves to provide expertise in the development and use of reagents for genetic perturbation, including RNAi, ORF overexpression and CRISPR-CAS9 gene editing platforms.
- Genetic Reagents: We generate and validate custom sgRNA vectors for CRISPR editing of individual genes. We provide assistance with creating CRISPR KO cells, as polyclonal pools or clonal KO lines. We also distribute individual lentiviral shRNA clones from the TRC are available as bacterial glycerol stocks, through our shRNA request system.
- Chemical Reagents: We have several libraries available for screening, see https://sites.duke.edu/functionalgenomics/libraries/compound-libraries/ for more details.
- Genome-wide screens: The Functional Genomics Shared Resource has the infrastructure necessary to conduct high-throughput screens, providing automated liquid handlers, cell platers and plate washers. For plate-based quantitative assays, we have the CellInsight CX5 high-content screening system, as well as multimodal plate readers. Screens can also be performed in pooled format using a selection-based strategy with any of our CRISPR or shRNA libraries. We can also assemble and screen custom CRISPR libraries.
- Phenotypic assays: The facility offers high-content screening with the CX5 HCS system, as well as sensitive signal detection with two high-performance multimodal plate readers. All systems are equipped with plate stackers and barcode readers for walk-away functionality and true high-throughput processing.