shRNA Libraries (human and mouse)
The Viral Vector Facility obtained the human and mouse Mission shRNA libraries of Sigma-Aldrich and The RNAi Consortium (TRC). You can use these libraries to study gene knockdown. Clones from these libraries can be requested for free for the purpose of research within the LUMC, but may not be distributed to external groups. shRNA constructs from these libraries can be used to produce lentiviruses.
- Complete human and mouse TRC1 and TRC1.5 libraries are available and the main part of TRC2 (all validated shRNA constructs, and all constructs of target genes which are not already present in TRC1 or TRC1.5).
- 129,500 human shRNA clones (20,000 unique target genes) and 118,000 mouse shRNA clones (21,200 unique target genes).
- TRC1 obtained in 2007; TRC1.5 in 2010-2013; and TRC2 in 2013.
- Bacterial glycerol stocks of all constructs are arrayed in 96-well plates (one clone per well).
- Plasmids are based on pLKO.1-puro vector. The backbones of TRC1 and TRC1.5 are identical. TRC2 has a minor modification; a wPRE element was included in the vector.
- We have control plasmids SHC-001 till SHC-009.
- Search for shRNA constructs: individual genes or multiple genes.
- Validated constructs were analysed by Sigma-Aldrich and gave a knockdown of at least 50% (high throughput qPCR). Validation data can be found on the Sigma-Aldrich website.
- To request shRNA constructs, please provide the accession/RefSeq number (NM_....../XM_......) of the target gene. If you don't want all available shRNA constructs of a target gene, please specify their TRC numbers (TRCN0000......).
- shRNA constructs are delivered as bacterial cultures (2 ml LB medium with ampicillin).
- This bacterial culture can be used to do a midi/maxi-prep and, if necessary, to generate lentivirus of it.