General description

This shRNA non-mammalian control was designed using our Turbo GFP sequence and may cause some knockdown of tGFP. For maximum knockdown of tGFP, please refer to SHC004, SHC004V, SHC004H, SHC204, or SHC204V.

The MISSION^® TRC2 Control Vector pLKO-puro Non-Target shRNA is a lentivirus plasmid vector. This vector is in the TRC2 pLKO-puro plasmid backbone, which contains the WPRE.

Ampicillin and puromycin antibiotic resistance genes provide selection in bacterial or mammalian cells respectively. In addition, self-inactivating replication incompetent viral particles can be produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids (SHP001). The TRC2 Non-Target shRNA Control Vector is provided as 10 μg of plasmid DNA in Tris-EDTA (TE) buffer at a concentration of 500 ng/μl.

Application

Small interfering RNAs (siRNAs) expressed from short hairpin RNAs (shRNAs) are a powerful way to mediate gene specific RNA interference (RNAi) in mammalian cells. The MISSION^® product line is based on a viral vector-based RNAi library against annotated mouse and human genes. shRNAs that generate siRNAs intracellularly are expressed from amphotropic lentivirus viral particles, allowing screening in a wide range of mammalian cell lines. In these cell lines, MISSION^® shRNA clones permit rapid, cost efficient loss-of-function and genetic interaction screens. The vector contains an shRNA insert that does not target human or mouse genes, making it useful as a negative control in experiments using the MISSION^® TRC2 shRNA library clones. This allows one to examine the effect of transfection of a short-hairpin on gene expression and interpret the knockdown effect seen with shRNA clones.

To see more application data, protocols, vector maps visit sigma.com/shrna.