miRNA Inhibitors

microRNA (miRNA) inhibitors block miRNA regulation of target gene expression. They are designed and optimized for miRNA loss of function study. LabOmics offers miArrest miRNA inhibitors as vector-based expression clones or synthetic oligonucleotides.

Vector-based expression clones are available in lentiviral and non-viral vectors. miRNA inhibitor clones 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

Advantages

• Full coverage

• All known human, mouse and rat miRNAs in the miRBase covered.

• Flexible delivery systems

• Lentiviral vectors allow efficient transduction and stable integration into the host genome of non-dividing and difficult to transfect cells target cells

  Non-viral vectors allow either transient or stable transfection

• Advanced and reliable design

• Expression cassettes have been designed using a proprietary algorithm and tested rigorously by carefully designed experiments

• Superior performance

• Superior potency, long lasting inhibition and extremely low cell toxicity compared to other chemically synthesized miRNA inhibitors (see table 1)

  Choices of H1 or U6 promoter allow constitutive expression of inhibitors in virtually all mammalian cell types.

• High-throughput compatible

• Quick and easy assay format

  High sample number compatible

Figure 1.

Data showing the effect of miArrest miRNA inhibitor clone in a dose-dependent manner

	Vector-Based Inhibitors	Synthetic 2'-OMe Inhibitors	LNA Inhibitors
Inhibition	+++++	++	++
Specificity	+++++	+++	+++
Stability	+++++	+	+++
Durability	Long term	Transient	Transient
Cell Toxicity	-	-	+
Delivery to resting and hard-to-transfect cells	+++++	-	-

Table 1. Comparison of vector-based inhibitors vs. synthesized miRNA inhibitors

Figure 2

Simplified mechanism of action for vector-based miRNA inhibitors

miRNA inhibitor constructs bind specifically to their target miRNA upon transduction into cells. The post-transcriptional processing causes formation of an entrapping structure that attracts and binds to two molecules of the intended miRNA.

This in turn prevents the binding of endogenous miRNA to target mRNA, thus inhibiting the effect of miRNA on target gene and facilitating enhanced target gene expression (figure 3).

A unique vector design ensures maximum inhibition with specificity and long-lasting stability while keeping off-target effects to a minimum.

miRNA inhibitor vector features

Vector	Reporter gene	Selection marker	Promoter	Type
pEZX-AM01	mCherry	Puromycin	H1	Mammalian
pEZX-AM02	mCherry	Puromycin	U6	Mammalian
pEZX-AM03	mCherry	Hygromycin	H1	HIV
pEZX-AM04	mCherry	Hygromycin	U6	HIV