The RNAi Consortium (TRC) shRNA human genome library clones are available as plasmids in E.Coli that can be used for generating lentivirus containing shRNA hairpin sequences that are able to knockdown the expression of specific genes in human cells.  There are about 118,000 clones covering ~20,000 genes. Individual clones can be acquired streaked on LB amp+ plates for $10/clone from the CHEO Research Institute HTS lab.

Vectors

The TRC 1.0 and 1.5 clones are contained in the pLK0.1-puro vector (snapgene file). The TRC2-pLK0.1 vector is the same except for the addition of the WPRE (Woodchuck Hepatitis Post-Transcriptional Regulatory Element). This allows for enhanced expression of transgenes delivered by lentivirus.

These are 3rd generation vectors that can be used in level 2 hoods and virus can be generated with the three additional plasmids, pMDLg/pRRE (snapgene file), pRSV-rev (snapgene file), and pVSV-G (snapgene file). Virus production protocols can be found on the Broad Institute protocol resource page.

Searching for clones

To search for what clones are available in the library for your gene(s) of interest, upload a plain text file containing the official gene symbols (eg PLK1) on each line. The file must have a .txt file extension. A spreadsheet will be returned with all the clones available for your gene queries. There will also be a spreadsheet with the gene symbols not found in the database. For any genes not found, check for typos in the gene name or if the official HUGO name has changed. There are also 2 columns which denote whether the shRNA have been validated to 70-80% knockdown in the cell line listed. Email the spreadsheet with only the rows of the clones you wish to obtain to the CHEO RI HTS Lab.