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一般描述
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.
We are proud to offer two different IPTG inducible vectors for your research. The preferred inducible vector, pLKO_IPTG_3xLacO, contains three lac operon sequences (two in the U6 promoter and one 3′ of the promoter) affording both tight regulation and great gene silencing. Whereas, the pLKO_IPTG_1xLacO vector contains a single lac operon sequence in the U6 promoter, which allows for an advantage to shRNA expression, but looser control of the promoter when not induced.
The MISSION® 3X LacO Inducible Non-Target shRNA Control is a lentivirus plasmid vector. 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.
We are proud to offer two different IPTG inducible vectors for your research. The preferred inducible vector, pLKO_IPTG_3xLacO, contains three lac operon sequences (two in the U6 promoter and one 3′ of the promoter) affording both tight regulation and great gene silencing. Whereas, the pLKO_IPTG_1xLacO vector contains a single lac operon sequence in the U6 promoter, which allows for an advantage to shRNA expression, but looser control of the promoter when not induced.
The MISSION® 3X LacO Inducible Non-Target shRNA Control is a lentivirus plasmid vector. 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.
应用
To see more application data, protocols, vector maps visit sigma.com/shrna.
法律信息
使用本产品需遵守一个或多个许可协议。有关详细信息，请参阅http://sigmaaldrich.com/missionlicense。
MISSION is a registered trademark of Merck KGaA, Darmstadt, Germany
Sigma is a registered trademark of Sigma-Aldrich Co. LLC
WGK
WGK 1
闪点(°F)
Not applicable
闪点(°C)
Not applicable
法规信息
常规特殊物品
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商品
When shRNA is delivered using lentiviral vectors, the sequence encoding the shRNA is integrated into the genome and the knockdown effect is passed on to daughter cells, continuing gene silencing.
RNAi Consortium (TRC): Collaborative effort among academic labs and biotech/pharma institutes advancing RNA interference research.
相关内容
Explore the benefits of IPTG-inducible vectors through performance data. These vectors offer regulated expression, which is important when studying essential and lethal genes.
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