Research and Development on Lentivectors for:
Lentivectors are relatively easy to produce on a small scale, by transient transfection as shown in this diagram.
The detailed recipes are at this link
On campus, we provide all the basic plasmids:
All these plasmids can also be obtained from Addgene.
Since 2005, we have developed a whole set of lentivectors containing the Gateway Recombination Cloning system. They can all be obtained from Addgene.
For single gene expression, you can choose your promoter and your gene of interest (GOI). You just need the Lenti-DEST platform (pCLX family), the pDONRP4P1R (to clone your promoter) and the pDONR221 (to clone your GOI). You run a LR reaction with the 3 plasmids, et voilà !
We also have bi-cistronic Lenti-DEST plasmids that allow for live sorting/tagging of transduced using GFP of mCherry living colors.
These lentis are especially suitable for in vivo applications.
We also have bi-cistronic Lenti-DEST plasmids containing selectable markers.
These lentis are more suitable for in vitro applications, such as the creation of mammalian cell lines.
And last but not least, we have developed a set of auto-inducible lentivectors, that are unique in terms of low level of basal (non induced) activity, versus high levels of induced activity.
They are called the Polyswitch Lentis and are described in this article.
Every detail has been optimized in these lentis, from the TET promoter (pTF), to the rtTAm2 transactivator and the 2A peptide for multiple protein expression from a single transcript.
These lentis are the best you can have if you want to remote-control the expression of a gene that would have an undesired effect even at low levels.
We have a version with the BSD selectable marker, allowing to generate cell lines and then induce the gene of interest at will.
We also have a version with GFP, allowing to track live transduced cells, and follow in these cells the effect of gene induction.
|Lentis Titration by FACS|
|Lentis Titration by qPCR|
|Design of efficient miRNAs|
|BSL-2 Emergency Procedures|
|BSL-2 Decontamination Procedures|
|Decontamination of Liquid Nitrogen Tanks|