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Concentration:
500 ng/μL in TE buffer; DNA (10μg of plasmid DNA)
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General description
Sigma® is proud to offer IPTG-inducible vectors as the latest development from our continued partnership in TRC.
The pLKO vector has been redesigned to contain a LacI (repressor) and a modified human U6 shRNA promoter with LacO (operator) sequences. In the absence of IPTG (isopropyl-Β-D-thio-galactoside), an analogue of lactose, LacI binds to LacO preventing expression of the shRNA. When IPTG is present, the allosteric LacI repressor changes conformation, releasing itself from lacO modified human U6 promoter, and subsequently allows expression of the shRNA.
The vector contains an shRNA insert that does not target any known genes, making it useful as a negative control in experiments using the MISSION inducible shRNA library clones. This allows one to examine the effect of transfection of a short-hairpin on gene expression and interpret the knockdown effect seen with shRNA clones. Ampicillin and puromycin antibiotic resistance genes provide selection in bacterial or mammalian cells respectively. In addition, self-inactivating replication incompetent viral particles can be produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids. The 1X LacO Inducible Non-Target shRNA Control is provided as 10 μg of plasmid DNA in Tris-EDTA (TE) buffer at a concentration of 500 ng/μl.
The pLKO vector has been redesigned to contain a LacI (repressor) and a modified human U6 shRNA promoter with LacO (operator) sequences. In the absence of IPTG (isopropyl-Β-D-thio-galactoside), an analogue of lactose, LacI binds to LacO preventing expression of the shRNA. When IPTG is present, the allosteric LacI repressor changes conformation, releasing itself from lacO modified human U6 promoter, and subsequently allows expression of the shRNA.
The vector contains an shRNA insert that does not target any known genes, making it useful as a negative control in experiments using the MISSION inducible shRNA library clones. This allows one to examine the effect of transfection of a short-hairpin on gene expression and interpret the knockdown effect seen with shRNA clones. Ampicillin and puromycin antibiotic resistance genes provide selection in bacterial or mammalian cells respectively. In addition, self-inactivating replication incompetent viral particles can be produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids. The 1X LacO Inducible Non-Target shRNA Control is provided as 10 μg of plasmid DNA in Tris-EDTA (TE) buffer at a concentration of 500 ng/μl.
Application
To see more application data, protocols, vector maps visit sigma.com/shrna.
Legal Information
Use of this product is subject to one or more license agreements. For details, please see http://sigmaaldrich.com/missionlicense.
MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany
Sigma is a registered trademark of Sigma-Aldrich Co. LLC
Storage Class
10 - Combustible liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
Regulatory Information
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Articles
RNAi Consortium (TRC): Collaborative effort among academic labs and biotech/pharma institutes advancing RNA interference research.
Related Content
Explore the benefits of IPTG-inducible vectors through performance data. These vectors offer regulated expression, which is important when studying essential and lethal genes.
Instructions
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