GeneCopoeia miTarget miRNA target sequence expression clones

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GeneCopoeia offers genome-wide microRNA 3′ UTR target constructs in mammalian expression vector systems with luciferase reporter genes for functional validation experiments.

With the target clone collections, all 3′ UTR sequences (inserted downstream of coding sequences) were obtained from public domain gene sequence databases. Expression cassettes are inserted downstream of the firefly or Gaussia luciferase reporter genes which are controlled by the SV40 promoter for expression in mammalian cells.

The Renilla luciferase or alkaline phosphatase genes can be used as a tracking indicator for successful transfection and expression of the miRNA constructs in target cells.

The regulatory effect of a particular miRNA on its potential target is assessed with a functional assay for the firefly or Gaussia luciferase.

Expression plasmids transcribe a chimeric mRNA consisting of the firefly or Gaussia luciferase coding sequence and a 3′ UTR target sequence.

Luciferase expression is regulated by binding of the targeting miRNA to the 3′ UTR target sequence and luciferase activity is quantified with a colorimetric assay.

Also available are the miArrest miRNA inhibitor clones which bind specifically to their target miRNAs, allowing transient as well as stable suppression of the target gene. The choice of H1 or U6 promoter allows constitutive expression of inhibitors in virtually all types of mammalian cells. All known human, mouse and rat miRNAs in miRBase are available.