The RNAi experts at NCATS employ scientific resources that include a variety of commercially available siRNA screening libraries; tools for data tracking, storage and analysis; and a fully automated, large-scale screening platform.
Screening Libraries
- Ambion Silencer Select Human Genome-Wide siRNA library targeting ~22,000 genes with three individual siRNAs per gene
- Ambion Silencer Mouse Genome-Wide siRNA library targeting ~17,000 genes with three individual siRNAs per gene
- Dharmacon siGENOME and Dharmacon ON-TARGETplus Human Genome-Wide siRNA libraries (Thermo Scientific), with pools of four siRNAs targeted against each of ~16,000 genes in the library; “druggable genome” subsets included for focused screening of potentially druggable targets
- microRNA mimic and inhibitor libraries
Software and Analysis Tools
- RNAi Data Viewer
This tool allows users to view plates, check stats, screen data and perform a variety of analyses. Access user data (restricted).
Assays
Simple phenotypes
Examples: viability, cytotoxicity
Assays that explore one aspect of morphology or function/loss of function.
Pathway reporter assays
Examples: luciferase, beta-lactamase
Assays using a specific read-out mechanism to “report” data, or “reporter assays.”
Complex phenotypes
Examples: high-content imaging, cell cycle, translocation
Assays that explore more than one aspect of morphology or function/loss of function, usually a process with two or more steps.
Robotic Platform
- Liquid handlers, washers and dispensers
- EnVision Multilabel Reader (PerkinElmer)
- Measures luminescence, fluorescence, fluorescence polarization, absorbance and time-resolved fluorescence
- ImageXpress Micro XL high-content imager (Molecular Devices)
- Automated microscope used for the acquisition and analysis of cell-based images in relatively high throughput
Schematic illustration of the RNAi robotic platform.
Photograph of the RNAi robotic platform.