lenti-shrna

Lenti-ONE shRNA

From €250.00

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.

 

shRNA vector map

 

immunochemistry-increasing inhibition of GFAP

western-increasing inhibition of GFAP

Vector doses are expressed in ng of p24; NT: Not Transduced. Results provided by GEG Tech R&D