The facility will provide the human genome siRNA library, perform the robotic spotting of the library in 384 well plates, guidance in screen development, support of instrumentation for screen development and implementation of primary and secondary screens.
Assay development and implementation of a screen can be performed in either a 384 well format using the Pherastar FS multifunctional plate reader and Cellomics high content imaging microscope. Secondary screens will be supported for validation of hits using single siRNAs, lentiviral shRNA clones, cell line overlap and high content imaging. The facility supports essentially any cell based screen for siRNA analysis including reporter genes, GFP/CFP/RFP readouts (examples: virus secretion/infection) viability (example: mix and measure Cell TiterGlo Luminescent Cell Viability Assay), mitotic arrest, complex phenotypes measured by cell morphology, fluorescent markers, immunostaining, etc.
Once a screen is performed, “hits” will be validated with the siRNA primary screen. Repeat activity in the primary screen will allow siRNAs to be used in a secondary screen.
Analysis of data will be performed using statistical methods for analysis of high-throughput RNA interference screens developed and supported by the RNAi Global Informatics Workgroup.
Brian Golitz, Director, provides data management and analysis for all screens performed in the facility. Brian will aid users in developing their screens, use of instrumentation and data analysis. Email: firstname.lastname@example.org
Dr. Gary Johnson will discuss siRNA screen proposals. Email: email@example.com
Noah Sciaky provides data analysis of HC imaging screens, when needed. Noah will work directly with users of the facility in developing HC imaging screens, implementation and analysis. Email: firstname.lastname@example.org