MISSION® TRC2 pLKO.5-puro-CMV-TurboGFP™ Positive Control Transduction Particles

Description

• General description: The MISSION TRC2 TurboGFP Control Transduction Particles are a critical positive control to monitor transduction efficiency. This control is produced from the sequence-verified lentiviral plasmid, TRC2 pLKO-puro TurboGFP (SHC203). This vector is in the TRC2 pLKO-puro plasmid backbone, which contains the WPRE. This construct aids in interpretation of experimental design and results by providing a green fluorescent protein marker for monitoring success in transduction of your cells of interest. TurboGFP is an improved variant of the green fluorescent protein copGFP cloned from the copepoda Pontellina plumata. 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.
• 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.
• Legal Information: MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany