High Throughput Screening
High-throughput screening (HTS) is a powerful tool for discovering novel targets or lead compounds as well as gaining mechanistic insight into cellular and molecular processes. Our facility provides access to expertise, reagents and infrastructure for high-throughput screening of genetic and chemical libraries. Facility personnel are available to support investigators through the entire screening process with both technical guidance and hands-on assistance with experiments. We assist in all steps of the screening process including screen design, assay development, pre-screen validation, screening, and data analysis. We encourage you to come talk to us early in the process to learn about the different screening formats and resources available for your project.
human ORF screening
We have a library of 600 lentiviral Open Reading Frame (ORF) expression vectors covering the human kinome from the CCSB-Broad ORF collection#.These can be used in arrayed or pooled screens. We also have the Sigma Mission TRC3 ORFeome-wide library of lentiviral human ORFs with barcodes for pooled screening.
#Johannessen, C et al, COT drives resistance to RAF inhibition through MAP kinase reactivation, Nature2010.
#Yang X et al, A public genome-scale lentiviral expression library of human ORFs, Nat Methods 2011.
Compound screening
We provide infrastructure, reagents and expertise to support high-throughput screens or reformatting of chemical libraries. We maintain chemical screening collections that can be used for drug discovery, chemical genetics, drug repurposing and combination studies. Currently available compound libraries include:
- Selleck FDA-approved drugs (978 compounds)
- Selleck Bioactives (2100 compounds)
- NCI Diversity Set V* (https://dtp.nci.nih.gov/organization/dscb/obtaining/available_plates.htm)
- NCI Approved Oncology Drugs Set VI* (https://dtp.nci.nih.gov/organization/dscb/obtaining/available_plates.htm)
- NCI Natural Products Set III* (https://dtp.nci.nih.gov/organization/dscb/obtaining/available_plates.htm)
- TimTec ActiGlobe 50K Diversity Set (http://www.timtec.net/actiglobe-50k.html)
- TimTec Natural Products Library 720 (http://www.timtec.net/natural-compound-library.html)
*For more information, please visit https://dtp.nci.nih.gov/organization/dscb/obtaining/available_plates.htm
CRISPR sgRNA libraries for pooled screening
We have several human and mouse genome and kinome lentiviral sgRNA libraries available for pooled loss- or gain-of-function screening. We also provide custom pooled library assembly and screening services. Please send inquiries to rnai@duke.edu to request more information on options for CRISPR sgRNA screening.
siRNA screening
Whole-genome siRNA screening: We currently offer screening of the human genome utilizing the Qiagen Human genome siRNA library. The library targets approximately ~22,000 genes with four unique siRNAs per gene, which can be screened as invidual or pooled siRNAs.
Sub-genome siRNA screening: The Qiagen Human genome siRNA library is also available in sub-genomic libraries targeting druggable genes, kinases, phosphatases,or GPCRs.
Custom siRNA sets: We can also work with investigators to design, order and screen custom designed sets of siRNAs against specific genes of interest.
shRNA screening
Custom lentiviral shRNA gene sets: We can also work with investigators to assemble custom designed gene sets from the TRC1 human lentiviral shRNA collection or siRNAs for screens in pooled or arrayed format.
