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miRNA Research Tools

Elucidate miRNA Function with MISSION® Lenti microRNA Inhibitors

 MicroRNA Inhibition in a Lentiviral Plasmid Vector

Inhibition of microRNAs is essential to studying their function. Sigma® Life Science offers a collection of individual microRNA inhibitors which are designed using a proprietary algorithm based upon the Tough Decoy (TuD) design.1 Each miRNA inhibitor construct has been cloned and sequence verified to ensure a match to the target miRNA.

 Benefits of Lenti miRNA Inhibitors

  • Allows for potent inhibition of desired miRNA
  • Lentiviral format allows for efficient delivery of the inhibitor into a wide variety of cell types
  • Enables long-term inhibition without repeat transfections

Lenti miRNA Inhibitor Design

Figure 1. Lentiviral miRNA Inhibitor Expression Regulates miRNA Function
Expression of the miRNA inhibitors is driven by the hU6 promotor upon genomic integration of the lentiviral transfer vector into the host cell post-transduction. miRNA inhibitors are able to competitively bind specific miRNAs and prevent them from regulating their endogenous targets.

 Ordering Information

Browse for your miRNA inhibitor by clicking the button below.

To place a custom MISSION Lenti miRNA Inhibitor order, please contact your local Sigma sales representative or click below and we will put you in touch with a sales rep who will place the order.


 Application Data

Validation of Lenti miRNA Inhibitors by Dual Luciferase Assay

Figure 2. Dual Luciferase Assay: HeLa Lenti miRNA Inhibitor Cell Lines
HeLa cells were stably transduced with lentivirus harboring the miRNA inhibitors indicated. Stable pools were cultured for a minimum of two weeks, and were then transfected with dual luciferase reporter constructs corresponding to the miRNA of interest. Ratios of renilla:firefly luciferase luminescence were calculated for each cell line tested, and compared to control inhibitor cells. An increase in Renilla:firefly luminescence in inhibitor-expressing cells relative to controls cells indicates functionality of the inhibitor.

Inhibition of miR-206 Upregulates Notch3 Expression

Figure 3. qRT-PCR of Notch3 in HeLa Lenti miRNA Inhibitor Cell Lines
miR-206 has been shown to regulate expression of Notch3 in HeLa cells. To confirm the miR-206 inhibitor can alter the actions of miR-206 with respect to its endogenous downstream targets, qRT-PCR was performed on total RNA preparations from HeLa cells stably expressing either the miR-206 inhibitor or control inhibitor. Expression of Notch3 increased by 78% relative to control cells, indicating efficient inhibition of miR-206 in this cell line.

Higher MOI may Increase miRNA Inhibition

Figure 4. Dual Luciferase Assay for miR-21 in HepG2 Cells
Increased multiplicity of infection (MOI) can be used to increase miRNA inhibition. HepG2 cells were stably transduced with control Lenti miRNA inhibitor or miR-21 inhibitor at varied MOI. Stable pools were transfected with the dual luciferase reporter vector for miR-21, and luciferase ratios were calculated accordingly. Results indicate an increased MOI of Lenti inhibitor can be used to increase inhibition of a specific miRNA, as the Renilla:firefly luminescence increases with higher MOI.

Inhibition of miR-21 Upregulates RHOB Expression

Figure 5. qRT-PCR of RHOB in HepG2 Lenti miRNA Inhibitor Cell Lines
miR-21 has been shown to regulate the expression of RHOB in hepatocytes. To further validate the miR-21 inhibitor, qRT-PCR was performed on total RNA preparations from HepG2 cells stably expressing either the miR-21 inhibitor or control inhibitor. Expression of RHOB increased by 55% relative to control cells, indicating efficient inhibition of miR-21 in this cell line. Nearly identical results were obtained for this same target and inhibitor combination in HeLa cells (data not shown).


Reference

  1. Haraguchi, T., et al., Vectors expressing efficient RNA decoys achieve the long-term suppression of specific microRNA activity in mammalian cells. Nucleic Acids Res., 37, (2009).