RNA interference is one of the many applications where lentiviral vectors can be used to significantly improve a project. Lenti-ONE shRNA carries the sequence of your shRNA of interest, driven by the U6 promoter.
Learn more about RNA interference
Lenti-ONE shRNA is a powerful tool to improve your vector-based RNAi experiments. Gene silencing efficiency depends on both the design and the level of expression of the shRNA. GEG Tech has a strong know-how in these fields and offers lentiviral vectors highly efficient in vitro as well as in vivo. Thanks to stringent QCs processes, GEG Tech guaranties a true titer superior to 1E9TU/mL for shRNA vectors with the standard vector configurations (1-3 shRNA combined with CMV-GFP reporter), and reaches this level of titer for almost all other cases. To perfectly fit your projects, Lenti-ONE shRNA can be combined with several vector options, such as:
- Different promoters to adapt the level of expression according to your experimental context
- Several choices of envelopes to target specific cell populations
- On/Off switch to control the shRNA expression
- Expression cassette designs allowing multiple shRNA expression
- Addition of reporters such as GFP or Neomycin to monitor your shRNA expression
- ANCH/OR system to live-track your Lenti-ONE vector in living cells
- Integrating or non-integrating Lenti-ONE versions to induce stable or transient expression
We also provides strong support to help you with your project all along the way. Our support team can advise on the design in order to find the best vector configuration for your project, and help you use your Lenti-ONE shRNA vector(s) to take full advantage of its potential and make your project successful.
Examples of Applications
Inhibition of endogenous GFAP expression using Lenti-ONE™ shRNA
Here is an example of RNA interference application using lentiviral vectorization. We have designed several shRNA targeting the swine GFAP sequence and selected the most efficient one through a rapid screening test, as illustrated in the above video. The selected shRNA sequence has been introduced into a lentiviral vector backbone. Results shown below demonstrate the efficiency of Lenti-ONE™ shRNA-GFAPsw to inhibit endogenous expression of GFAP in vitro in primary swine astrocytes.
Primary swine astrocytes have been transduced in vitro with increasing doses of Lenti-ONE™ shRNA-GFAPsw. These vectors express a short hairpin inducing an RNA interference mechanism targeting the endogenous swine GFAP mRNA under the control of a U6 promoter, along with a CMV-GFP cassette allowing the monitoring of transduction efficiency. Cells were fixed 7 days after transduction and both GFP and GFAP expression were analysed through immunochemistry. As you can see below, increasing doses of Lenti-ONE™ shRNA-GFAPsw induce increasing expression on GFP and reduced expression of GFAP. Inhibition of GFAP expression has been further confirmed with Western Blot analysis.
Vector doses are expressed in ng of p24; NT: Not Transduced. Results provided by GEG Tech R&D