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LightSwitch™ miRNA Mimics and Inhibitors

study how miRNA-3´UTR interactions impact gene regulation

MicroRNAs (miRNAs) are important post-transcriptional regulators that are thought to repress expression by targeting the 3´UTR of mRNA transcripts. The interaction of a miRNA with a 3´UTR results in either the degradation of the target transcript or inhibition of translation.

LightSwitch™ miRNA Mimics are transfected into cells to study the effects of over-expression of a miRNA of interest. In contrast, LightSwitch miRNA Inhibitors are used to knock down the expression of endogenous human miRNAs in living cells.

All LightSwitch miRNA products have been optimized for use with our collections of over 12,000 LightSwitch 3´UTR Reporter constructs and over 900 optimized LightSwitch Synthetic miRNA Target Reporter constructs. The combination of LightSwitch miRNA Mimics and Inhibitors with our 3´UTR reporter clones provides a fully integrated system for validating the 3´UTR targets of your miRNAs of interest. In addition, we offer Custom miRNA Target Validation Services in which our scientists can validate the targets of your miRNAs of interest using our high-throughput, cell-based assay.

IMPORTANT:  Because all LightSwitch reporter constructs contain the optimized RenSP luciferase gene, you MUST use our LightSwitch Luciferase Assay Reagents to obtain optimal results. (Other luciferase assay reagents are not compatible with RenSP.)

Find miRNA Mimics & Inhibitors by clicking one of the groups below:

 

miRNA-3´UTR interactions regulate gene expression

Graph demonstrating knock-down of 3'UTR targets by miR-155 mimic
Figure 1: Knock-down of 3´UTR targets in the presence of miR-155 Mimic.

Four LightSwitch 3´UTR Reporter GoClones containing validated targets of miR-155 and two negative control vectors (Random 3´UTR Control 1 and pLightSwitch_3UTR with No Insert) were co-transfected with 50 nM of miRNA mimic or a non-targeting control miRNA in K-562 cells. The knockdown of each vector was calculated as the log2 ratio of miR-155 signal divided by the non-targeting control signal. The four known 3´UTR targets of miR-155 show a strong knockdown compared to the two negative control vectors.

The data above demonstrates the knock-down in luciferase signal from LightSwitch 3´UTR Reporter GoClone constructs that contain validated targets of miR-155 when co-transfected with miR-155 Mimic relative to co-transfection with a non-targeting control miRNA.

There are approximately 1000 miRNAs in the human genome; they range in length from 21-25 nucleotides. miRNAs regulate gene expression by binding to the 3-prime Untranslated Region (3´UTR) of messenger RNA transcripts (mRNAs). This interaction can both affect the rate of transcript degradation as well as the efficiency of translation. Both mechanisms result in a reduction in the amount of protein produced from the mRNA.

LightSwitch miRNA Mimics can be used to study the over-expression of a miRNA in a particular cell line, whereas LightSwitch miRNA Inhibitors can be used to study the loss of function of a particular miRNA in a cell line where the miRNA is endogenously expressed.

LightSwitch miRNA Mimics are chemically optimized synthetic double-stranded RNAs that act as functional equivalents to endogenous human miRNAs. They can be transfected into cells to study the effects of over-expression of a miRNA of interest. They can also be co-transfected with LightSwitch 3´UTR Reporter GoClone constructs to validate the 3´UTR targets of your miRNAs of interest (Figure 1).

LightSwitch miRNA Inhibitors are chemically optimized synthetic single-stranded RNAs that are used to knock down the expression of endogenous human miRNAs in living cells. They can be transfected into cells to study the effects of the loss of function of a miRNA of interest and can also be co-transfected with 3´UTR Reporter GoClone constructs for 3´UTR target validation.

All LightSwitch miRNA products are based on miRNA sequences deposited in the miRBase database, and they have been optimized for use with our collection of over 12,000 LightSwitch 3´UTR Reporter constructs and our over 900 LightSwitch Synthetic miRNA Target Reporter constructs.

Contents & Storage

LightSwitch miRNA Mimics and Inhibitors are shipped as lyophilized pellets. The lyophilized miRNA products are stable at -20°C. After resuspension, the products can be stored at -80°C. For maximum stability, minimize the number of freeze-thaw cycles of the products in solution. All reagents are guaranteed stable for 6 months when stored properly.