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Key Documents

SHC201V

Sigma-Aldrich

MISSION® TRC2 pLKO.5-puro Empty Vector Control Transduction Particles puro

Contains no shRNA insert

Synonym(s):

MISSION®, MISSION® Control Transduction Particles

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About This Item

UNSPSC Code:
41106609
NACRES:
NA.51

Quality Level

product line

MISSION®

concentration

≥1x106 VP/ml (via p24 assay)

shipped in

dry ice

storage temp.

−70°C

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General description

The MISSION TRC2 pLKO-puro Control Transduction Particles are produced from the sequence-verified lentiviral plasmid, TRC2 pLKO-puro Control Vector (SHC201). This vector is in the TRC2 pLKO-puro plasmid backbone, which contains the WPRE. The vector does not contain an shRNA insert and is useful as a negative control in experiments using the TRC2 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.
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

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.
To see more application data, protocols, vector maps visit sigma.com/shrna.
capture ELISA: 106 TU/mL using p24

Legal Information

MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable


Certificates of Analysis (COA)

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R Zufferey et al.
Journal of virology, 73(4), 2886-2892 (1999-03-12)
The expression of genes delivered by retroviral vectors is often inefficient, a potential obstacle for their widespread use in human gene therapy. Here, we explored the possibility that the posttranscriptional regulatory element of woodchuck hepatitis virus (WPRE) might help resolve
J E Donello et al.
Journal of virology, 72(6), 5085-5092 (1998-05-30)
The hepatitis B virus posttranscriptional regulatory element (HBVPRE) is a cis-acting RNA element that partially overlaps with enhancer I and is required for the cytoplasmic accumulation of HBV surface RNAs. We find that the closely related woodchuck hepatitis virus (WHV)

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