- Clone miRNA targets and evaluate miRNA regulation
- Screen putative miRNA target sequences
- Includes both reporter and control vectors
The pMIR-REPORT™ miRNA Expression Reporter Vector System provides accurate, quantitative, in-cell measurement of miRNA expression. This validated reporter system contains two mammalian expression vectors (see figure at right). The pMIR-REPORT™ Luciferase miRNA Expression Reporter Vector contains firefly luciferase under the control of a mammalian promoter/terminator system, with a miRNA target cloning region downstream of the luciferase translation sequence. This vector is optimized for cloning of miRNA targets and evaluation of miRNA regulation. A second vector, pMIR-REPORT™ Beta-galactosidase Reporter Control Vector, is provided for normalization of transfection efficiency.
Applications
pMIR-REPORT Luciferase is designed for the cloning and testing of putative miRNA binding sites. pMIR-REPORT Luciferase can be transfected into mammalian cells to evaluate endogenous miRNA expression, or used to evaluate the up- or down-regulation resulting from the transfection of Pre-miR™ miRNA Molecules or Anti-miR™ miRNA Inhibitor Molecules respectively. pMIR-REPORT Luciferase can also be used as a sequence screening tool to identify miRNA targets or screen libraries of Pre-miR miRNA Molecules to identify genes that regulate expression.
Product Format
The pMIR-REPORT miRNA Expression Reporter Vector System includes two reporter vectors (pMIR-REPORT Luciferase miRNA Expression Reporter Vector and pMIR-REPORT Beta-gal Reporter Control Vector) as supercoiled plasmids pre-transformed into E. coli and supplied as a glycerol stock.
pMIR-REPORT Luciferase utilizes the powerful CMV promoter to drive high-level expression of firefly luciferase in mammalian cells. A miRNA target multiple cloning site follows the luciferase gene and is itself followed by a SV40 polyA region. The vector utilizes puromycin for selection in cell culture and a ColE1 Ori/Ampicillin-resistance gene for maintenance in E. coli.
The pMIR-REPORT Beta-gal Control Vector is similar to pMIR-REPORT Luciferase, but has a beta-gal rather than a luciferase expression reporter and is intended for use in the normalization of transfection efficiency.
Ambion scientists describe their use of this product in the following publication:
Cheng AM, Byrom MW, Shelton J, Ford LP. Antisense inhibition of human miRNAs and indications for an involvement of miRNA in cell growth and apoptosis. Nucleic Acids Res 33(4): 1290-7. [read]
Trademarks, Patents, Licensing
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