SHC002V
MISSION® pLKO.1-puro Non-Mammalian shRNA Control Transduction Particles
Targets no known mammalian genes
Synonym(s):
MISSION®, MISSION® Control Transduction Particles
About This Item
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Quality Level
100
200
product line
MISSION®
concentration
≥1x106 VP/ml (via p24 assay)
technique(s)
capture ELISA: 106 TU/mL using p24
shipped in
dry ice
storage temp.
−70°C
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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 an shRNA lentiviral non-target control plasmid. It is useful as a negative control in experiments with the MISSION shRNA target sets.
Unlike murine-based MMLV or MSCV retroviral systems, lentiviral-based particles permit efficient infection and integration of the specific shRNA construct into differentiated and non-dividing cells, such as neurons and dendritic cells,1 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.2-3
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.4 The lentiviral transduction particles are titered via a p24 antigen ELISA assay and pg/ml of p24 are then converted to transducing units per ml using a conversion factor. The conversion can be viewed at: www.tronolab.com.
To see more application data, protocols, vector maps visit sigma.com/shrna.
Application
Legal Information
recommended
WGK
WGK 3
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
Certificates of Analysis (COA)
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Articles
Methods for lentiviral transduction of Jurkat cells were compared. Spinoculation was compared with overnight incubation with polybrene (hexadimethrine bromide) and fibronection-coated plates.
Protocols
Detailed procedure for how to perform a lentiviral transduction of MISSION shRNA lentiviral particles to achieve a stable long term silencing and phenotypic change.
This detailed procedure allows you to transduce Mouse Embryonic Fibroblasts (MEF) using MISSION ExpressMag Super Magnetic Kit.
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