SHC002V
MISSION® pLKO.1-puro Non-Mammalian shRNA Control Transduction Particles
Targets no known mammalian genes
Synonym(e):
MISSION®, MISSION® Control Transduction Particles
About This Item
Empfohlene Produkte
Qualitätsniveau
100
200
Produktlinie
MISSION®
Konzentration
≥1x106 VP/ml (via p24 assay)
Methode(n)
capture ELISA: 106 TU/mL using p24
Versandbedingung
dry ice
Lagertemp.
−70°C
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Allgemeine Beschreibung
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.
Anwendung
Rechtliche Hinweise
Empfehlung
Lagerklassenschlüssel
12 - Non Combustible Liquids
WGK
WGK 3
Flammpunkt (°F)
Not applicable
Flammpunkt (°C)
Not applicable
Persönliche Schutzausrüstung
Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)
Analysenzertifikate (COA)
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Artikel
Methods for lentiviral transduction of Jurkat cells were compared. Spinoculation was compared with overnight incubation with polybrene (hexadimethrine bromide) and fibronection-coated plates.
Protokolle
Detailliertes Verfahren zur Durchführung einer lentiviralen Transduktion von MISSION shRNA lentiviralen Partikeln, um eine stabile, langfristige Stilllegung und Phänotypveränderung zu erreichen.
This detailed procedure allows you to transduce Mouse Embryonic Fibroblasts (MEF) using MISSION ExpressMag Super Magnetic Kit.
Detailed procedure for how to perform a lentiviral transduction of MISSION shRNA lentiviral particles to achieve a stable long term silencing and phenotypic change.
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