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microRNA Target Site Blockers

High-affinity LNA™-enhanced microRNA target site blockers (TSBs) for the study of single microRNA target sites. Ideal for determining which pathway is involved in the observed effects of microRNA inhibition.

  • Custom designed target site blockers for specific inhibition of microRNA targets for all microRNAs in miRBase
  • Sophisticated and innovative design
  • Superior high affinity regardless of target sequence
  • Unmatched high efficacy in vitro and in vivo
  • Unrivaled performance – LNA™ TSBs do not catalyze RNase H-dependent mRNA degradation which leads to higher protein expression
  • Efficient at very low concentrations – The high affinity of LNA™ means that the TSBs outcompete microRNAs for their target sites
  • Superior biological stability for long lasting antisense activity
  • Optimized in vivo design options with phoshorothioate backbone and low toxicity risk

What are target site blockers?

MicroRNA target site blockers are antisense oligonucleotides that bind to the microRNA target site of an mRNA thereby preventing microRNAs from gaining access to that site. This allows researchers to study the effects of a microRNA on a single target. In contrast, the phenotype observed when inhibiting a microRNA reflects the combined effects of that microRNA on all targets.


LNA™-enhanced target site blockers

The incorporation of LNA™ in the miRCURY LNA™ microRNA Target Site Blockers means that they will compete more effectively with microRNA/RISC complex for the microRNA target site (Figure 1). In addition, LNA™ distribution throughout the LNA™/DNA mixmer ensures that the antisense oligonucleotide does not catalyze RNase H-dependent degradation of the mRNA. As a result, the TSB will cause increased expression of the protein encoded by the targeted mRNA by preventing microRNA-mediated translational attenuation (Figure 2).


Coverage

miRCURY LNA™ microRNA Target Site Blockers are available in several different categories depending on application (Table 1).

Table 1
Overview of miRCURY LNA™ microRNA TArget Site Blockers. (Click to learn more)





Figure 1 LNA™-enhanced target site blockers compete effectively with RISC for microRNA binding site
LNA™-enhanced target site blockers compete effectively with RISC for microRNA binding site. (Click to learn more)



Figure 2 Target site blockers stimulate translation of specific mRNAs by masking the microRNA target sites.
Target site blockers stimulate translation of specific mRNAs by masking the microRNA target sites. (Click to learn more)



Unravel miRNA function with target site blockers

MicroRNAs typically regulate gene expression of multiple targets. Inhibition of the microRNA will result in derepression of all these targets. A phenotype observed upon microRNA inhibition is a composite result of derepression of several targets (Figure 1). However, often deregulation of a few of these targets will contribute significantly to the phenotype. Identification of these targets is important to understanding the function of the microRNA.

microRNA TSBs can be used to:
  • Determine which pathway is involved in the phenotype observed upon microRNA inhibition
  • Determine which microRNA/mRNA interactions are most important in a pathway containing several predicted targets.
Examples of TSB applications are described in Figure 2.

Figure 3 shows an example of where a target site blocker has been used to show how miR-199a-5p functions as a key effector of TGFβ signaling in lung fibroblasts.
Figure 1 microRNA Inhibitors vs. target site blockers.
microRNA Inhibitors vs. target site blockers. (Click to learn more)




Figure 2 Examples of TSB applications.
Examples of TSB applications. (Click to learn more)




Figure 3 The CAV1 target site blocker phenocopies  the miR-199a-5p inhibitor in many important aspects
The CAV1 target site blocker phenocopies the miR-199a-5p inhibitor in many important aspects. (Click to learn more)
Dr. Magali Taulan

Development of antisense therapy for cystic fibrosis


"In our hands, in addition to high affinity, LNA™ antisense oligonucleotides display excellent activity upon simple addition to the cell culture – without the need for transfection reagents – which is great advantage when working with primary cell cultures."

Dr. Magali Taulan is an associate professor at the University of Montpellier in France. Her lab has been using LNA™ antisense oligonucleotides that interfere with microRNAs to better understanding how expression of the gene (CFTR) is regulated. As a result, the study potentially identify new targets for treatment of CF.

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Dr. Virgine Mattot

MicroRNA function in endothelial cells


Dr. Mattot studies the roles played by microRNAs in endothelial cells during physiological and pathological processes such as angiogenesis or endothelium activation. The fact that her microRNA of interest had a predicted target gene which was previously uncharacterized was a major challenge. However through the use of specific target site blockers, it was possible to demonstrate that this unknown gene was associated with the phenotype observed when the microRNA was inhibited in endothelial cells.

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