I have recently used Sigma esiRNA against SAT1 gene, and I was very surprissed when I checked the transfections using Real Time PCR and discovered that, in trhee independent experiments, cells treated with siRNA for 24h showed more mRNA levels than non transfected cells; the universal negative control also induced expression although not to the same extent. I insist, this happened in three independent experiments.
Other member of my research group used Sigma esiRNA against another gene and he had a mRNA induction too. He also checked de PCR products in an agarose gel, and the bands showed that transfected cells had more mRNA than controls.
I wonder if someone has ever had some problems like this, and if there is any explanation and solution.
Thank you in advance for your help.
Sigma esiRNA induces target gene expression
1 reply to this topic
Posted 19 June 2011 - 08:19 PM
This is weired, though I have often heard that people see increase in gene expression while expecting RNAi. Can you confirm the increase in mRNA expression by Western? Why don't you just use a single siRNA for your target gene.