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SHC002V Sigma-Aldrich

MISSION® pLKO.1-puro Non-Mammalian shRNA Control Transduction Particles

Targets no known mammalian genes

Synonym: MISSION® TurboGFP® Control Transduction Particles

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Properties

Related Categories Cell Biology, Functional Genomics and RNAi, General Stem Cell Biology, MISSION Controls in Lentiviral Format, MISSION shRNA,
product line   MISSION®
application(s)   capture ELISA: 106 TU/mL using p24
shipped in   dry ice
storage temp.   −70°C

Description

General description

This shRNA non-mammalian control was designed using our Turbo GFP sequence and may cause some knockdown of tGFP. For maximum knockdown of tGFP, please refer to SHC004, SHC004V, SHC004H, SHC204, or SHC204V.

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.

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.

Application

MISSION® pLKO.1-puro Non-Mammalian shRNA Control Transduction Particles has been used as a negative control in ACSS2 (cytosolic acetyl-CoA synthetase) knock down study. It has also been used to study the effects of transduction.

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

Legal Information

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

Safety & Documentation

Safety Information

RIDADR 
UN 3245 9
WGK Germany 
3

Milli-Q® Water Purification Solutions
Protocols & Articles

Articles

Enhancing Lentiviral Transduction Efficiency

An experiment to directly compare three methods of lentiviral transduction of Jurkat cells was conducted in order to determine the method that yields the greatest transduction efficiency. Spinoculati...
Andrea Spencer
LSI Edition 25
Keywords: Apoptosis, Centrifugation, Diseases, Gene expression, Methods, Protocols, Transduction, Type, Vectors

Protocols

Lentiviral Transduction Protocol

Prepare mammalian cells so that they are growing exponentially and are no more than 70-80% confluent before transduction.
Keywords: Cell culture, Gene expression, Molecular biology, Transduction

Protocol for Transducing Mouse Embryonic Fibroblasts (MEF) using MISSION® ExpressMag™ Super Magnetic Kit®

0.1 Seed 15k MEF per well into 96-well plate. Utilize a final volume of 200 µl of media per well. (Note: for best data interpretation always include an untransduced well that is treated with puromyci...

Peer-Reviewed Papers
15

References

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SHC002 MISSION® pLKO.1-puro Non-Mammalian shRNA Control Plasmid DNA, Targets no known mammalian genes
SHC001V MISSION® pLKO.1-puro Empty Vector Control Transduction Particles, Contains no shRNA insert
SHC003 MISSION® pLKO.1-puro-CMV-TurboGFP Positive Control Plasmid DNA, Green fluorescent protein marker to monitor transduction efficiency
SHC003V MISSION® pLKO.1-puro-CMV-TurboGFP Positive Control Transduction Particles, Green fluorescent protein marker to monitor transduction efficiency
SHC004 MISSION® pLKO.1-puro TurboGFP shRNA Control Plasmid DNA, shRNA sequence targeting tGFP
SHC004V MISSION® pLKO.1-puro TurboGFP shRNA Control Transduction Particles, shRNA sequence targeting tGFP
SHM01 MISSION® ExpressMag® Super Magnetic Kit, Increases transduction efficiency Enhance your lentiviral transduction!
SHM02 MISSION® ExpressMag® 96-Well Magnetic Kit, Increases transduction efficiency Enhance your lentiviral transduction in 96-well plate format!

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