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 MISSION® 1x LacO Inducible TurboGFP™ shRNA Control Transduction Particles contain an shRNA sequence targeting TurboGFP. The TurboGFP shRNA Control Particles are useful as a positive knockdown control in experiments using the MISSION TurboGFP positive control vector or in cell lines expressing TurboGFP. TurboGFP is an improved variant of the green fluorescent protein copGFP cloned from copepoda Pontellina plumata.
The TurboGFP shRNA control transduction particles are produced from the sequence-verified lentiviral plasmid, pLKO.1-puro-TurboGFP shRNA (SHC004). Self-inactivating replication incompetent viral particles are produced in packaging cells (HEK293T) by co-transfection with compatible packaging plasmids. In addition, the Control 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. 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.
MISSION is a registered trademark of Sigma-Aldrich Co. LLC
Sigma is a registered trademark of Sigma-Aldrich Co. LLC
TurboGFP is a trademark of Evrogen Co.