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SHC002V

Sigma-Aldrich

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

Targets no known mammalian genes

Synonym(s):

MISSION®, MISSION® Control Transduction Particles

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About This Item

UNSPSC Code:
41106609
NACRES:
NA.51

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

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 Merck KGaA, Darmstadt, Germany

WGK

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

Certificates of Analysis (COA)

Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.

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P2X4 assembles with P2X7 and pannexin-1 in gingival epithelial cells and modulates ATP-induced reactive oxygen species production and inflammasome activation
Hung SC,et al.
PLoS ONE, 8(7), e70210-e70210 (2013)
Bin J, et al.
Cancer Letters, 342(1), 104-112 (2014)
Panu K Luukkonen et al.
Journal of hepatology, 67(1), 128-136 (2017-02-27)
Carriers of the transmembrane 6 superfamily member 2 E167K gene variant (TM6SF2 Liver biopsies were taken from subjects characterized with respect to the TM6SF2 genotype, serum and liver lipidome, gene expression and histology. In vitro, after TM6SF2 knockdown in HuH-7
David Gilot et al.
Nature cell biology, 19(11), 1348-1357 (2017-10-11)
Competition among RNAs to bind miRNA is proposed to influence biological systems. However, the role of this competition in disease onset is unclear. Here, we report that TYRP1 mRNA, in addition to encoding tyrosinase-related protein 1 (TYRP1), indirectly promotes cell
Tumor uptake of radiolabeled acetate reflects the expression of cytosolic acetyl-CoA synthetase: implications for the mechanism of acetate PET
Yoshii Y, et al.
Nuclear Medicine and Biology, 36(7), 771-777 (2009)

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

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.

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

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