High-Throughput Genome-Wide Screening

The Lenti-shRNA Library Core gives investigators at Cincinnati Children’s and UC access to cutting-edge high-throughput screening (HTS), allowing for non-biased discovery of novel genes that affect biologic pathways of interest. The screening process uses cell-based systems and viral-mediated transduction with shRNA libraries developed by the RNAi Consortium (TRC). The Lenti-shRNA Library Core utilizes TRC-developed genome-wide mouse and human libraries that together encode a total of 160,000 shRNA clones against 32,000 genes, and they have been designed for fully automated handling and production of lentivirus.  

The system is highly amendable for screening projects in cell-based 96-well and 384-well culture plate formats. Investigators have the option of ordering single genes through our “Order Your Gene” service, or can request whole-library screening. This technology holds tremendous power, and is ready to help investigators at Cincinnati Children’s and UC work toward breakthrough discoveries.

Lenti-shRNA Library Features:

  • (21mer) designed against NCBI REFSEQ
  • shRNA candidates initially scored for efficiency, specificity and spacing
  • Five candidates selected for every gene target (including a 3'UTR clone)
  • Human Library: 16,018 genes, 81,895 clones (1000-96 well plates)
  • Mouse Library: 15,959 genes, 77,713 clones (820-96 well plates)
  • 99.6 percent of genes have a minimum of four clones, many have more than five

The mouse and human libraries were purchased from Sigma-Aldrich, which now manages and holds the rights to the library (MISSION shRNA). Funds to purchase the library were provided by the Howard Hughes Medical Institute (Jeff Molkentin, investigator), the Cincinnati Children’s Heart Institute (Jeff Robbins, co-director) and the Department of Cancer Cell Biology at the University of Cincinnati.