Lenti-ONE shRNA – Lentiviral vector
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
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