The Mir-X Inducible miRNA Red & Green Systems exploit key elements of our tightly regulated and highly responsive Tet-On Advanced Expression System to provide on-demand, high-level expression of your selected microRNA and a bright red or green fluorescent protein marker.
How It Works…
Your microRNA sequence is embedded in the 3’ UTR of a marker protein mRNA, which is expressed from an inducible miRNA expression vector (pmRi-mCherry or pmRi-ZsGreen1). When cells expressing the Tet-On Advanced transactivator are transfected with the pmRi-mCherry or pmRi-ZsGreen1 vector, high levels of your inducible microRNA and either the mCherry or ZsGreen1 proteins are expressed in response to doxycycline (Dox). Once Dox is added to the cell culture medium, miRNA induction is very rapid and effective.
We used modified Metridia-secreted luciferase genes containing miRNA target sequences (RNAiMonitor) to measure the effects of miR-1 and miR-9 miRNA induction in MCF7 Tet-On Advanced cells. Inducing miR-1 and miR-9 expression in these cells with Dox resulted in >90% knockdown of luciferase activity compared to controls (see Images & Data tab).
Red and Green Complete Systems
The complete Mir-X Inducible miRNA Systems provide the pTet-On Advanced Vector for expressing the Dox-responsive transactivator; an inducible miRNA expression vector (either pmRi-mCherry or pmRi-ZsGreen1; also available separately); two linear selectable markers (hygromycin and puromycin) for cotransfecting with the pmRi vectors in order to establish stable cell lines; and tetracycline-free Tet System Approved FBS (serum) for ensuring the full activity of Tet-based expression systems.