MicroRNA Functional Analysis Workflow

Workflow Features

  • Gain- and loss-of-function phenotypes identify genes and processes regulated by miRNAs
  • Western blotting and/or real-time RT-PCR are used to validate miRNA targets
  • Target site interaction and impact on protein expression can be evaluated using reporter gene and immunodetection systems

How It Works

  • Ambion® Pre-miR™ miRNA Precursors and Anti-miR™ miRNA Inhibitors are designed to mimic or inhibit specific miRNAs for artificial upregulation and downregulation of target mRNA translation, respectively
  • miRNA targets are validated by quantitating target protein and/or messenger RNA levels in response to miRNA upregulation or downregulation
  • Western analysis is used to investigate the impact on protein expression
  • Reporter gene assays are used to study the interaction between miRNAs and their target sites