General description

The MISSION TRC2 Control Vector pLKO-puro is a lentivirus plasmid vector. This vector is in the TRC2 pLKO-puro plasmid backbone, which contains the WPRE. The vector does not contain an shRNA insert and is useful as a negative control in experiments using the TRC2 MISSION shRNA library clones. This allows one to examine the effect of transfection on gene expression and interpret the knockdown effect seen with shRNA clones.

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 pLKO-puro Control Vector is provided as 10 μg of plasmid DNA in Tris-EDTA (TE) buffer at a concentration of 500 ng/μl.

Application

MISSION^® TRC2 pLKO.5-puro Empty Vector Control Plasmid DNA has been used in:

CRISPR library generation
Viral constructs
Plasmid construction
CRISPR/Cas9-mediated knockouts.

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.

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