UNC Scientific Investigators:
The new LCCC RNAi High-throughput Screening (HTS) Facility is now officially open and provides the technology for the UNC scientific community to perform cell based screens to define the contribution of each gene in the human genome to specific phenotypes. RNAi mediated inhibition of gene expression is a powerful method for identifying genes and their cognate phenotype, whose loss of expression alters the cell phenotype. The RNAi HTS Facility manages a human genomic siRNA library from Dharmacon. The library is composed of ~84,500 siRNAs targeting 21,125 genes (Ref Seq database v5.0-8.0). The Services and Fees pages describe what the facility provides including technical support and the cost for the investigator to develop and implement a screen. A dedicated scientist from the investigator's laboratory will be required to work with facility personnel to develop and implement a screen. After validation of a primary screening assay, a genome-wide siRNA screen for all 21,125 genes will average 8 weeks of dedicated work in the facility. Targeting a specific class of genes such as protein kinases or ubiquitin ligases is an option and requires less time and resources (for example, the kinases represent 9 96 well plates whereas the genome is formatted in 231 plates). Additional time and investigator expenditures will be needed for validation of hits using the primary screen and then secondary screen assay.
Questions regarding the development of a siRNA screen should be addressed to Gary Johnson at glj@med.unc.edu. We strongly encourage UNC investigators to think about human cell-based genome siRNA screens in their research. This is a very powerful new experimental resource for UNC supported by the LCCC.
H. Shelley Earp
Gary L. Johnson
