MISSION^® pLKO.1-puro Empty Vector Control Transduction Particles

Application

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

General description

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 lentiviral transduction particles are produced from the sequence-verified lentiviral plasmid, pKLO.1-puro Control Vector. The vector does not contain an shRNA insert and is useful as a negative control in experiments using the MISSION shRNA library clones. This allows one to examine the effect of transduction on gene expression and interpret the knockdown effect seen with shRNA clones.

Unlike murine-based MMLV or MSCV retroviral systems, lentiviral-based particles permit efficient infection and integration of the construct into differentiated and non-dividing cells, such as neurons and dendritic cells, overcoming low transfection and integration difficulties when using these cell lines. Self-inactivating replication incompetent viral particles are produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids.

In addition, the lentiviral transduction particles are pseudotyped with an envelope G glycoprotein from Vesicular Stomatitis Virus (VSV-G), allowing transduction of a wide variety of mammalian cells. 200 μl of 10⁶ TU/ml (via p24 titering assay) lentiviral particles are provided as frozen stock.

When conducting experiments using MISSION^® shRNA clones, the proper controls should be a key element of your experimental design to allow for accurate interpretation of knockdown results. The MISSION Control Transduction Particles are a critical positive control to monitor transduction efficiency.
To see more application data, protocols, vector maps visit sigma.com/shrna.

Legal Information

MISSION is a registered trademark of Sigma-Aldrich Co. LLC