Takara Tetracycline-inducible miRNA

Takara Tetracycline-inducible miRNA (Mir-X Inducible miRNA Red and Green Systems)

The Mir-X inducible miRNA red and green systems exploit key elements of the Tet-On Advanced Inducible Expression System to provide controlled co-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 these vectors, 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 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 induction in MCF7 Tet-On Advanced cells.
Inducing miR-1 and miR-9 expression with Dox resulted in >90% knockdown of luciferase activity compared to controls.

Red and Green Complete Systems

The complete Mir-X inducible miRNA systems provide:

• pTet-On Advanced Vector for expressing the Dox-responsive transactivator
• Inducible miRNA expression vector (either pmRi-mCherry or pmRi-ZsGreen1, also available separately)
• Two selectable markers (hygromycin and puromycin) for cotransfection and stable cell line establishment
• Tetracycline-free Tet System Approved FBS (serum) for ensuring full activity of Tet-based expression systems

Overview

• Track your inducible miRNA expression with fluorescent protein markers
• High fold-induction and low basal expression

Applications

• Tetracycline-inducible miRNA expression in mammalian cells

Additional Product Information

Please refer to the product’s Certificate of Analysis for details on storage conditions, product components, and technical specifications.
The Kit Components List is available under the Documents tab.

Specifications