MicroRNA Sponge Service

AcceGen offers high-quality service for the construction of miRNA sponge vector clones to assist researchers worldwide in their miRNA studies. MicroRNAs (miRNAs) are short non-coding RNA molecules that regulate the expression of their target genes at the post-transcriptional level by binding to their respective mRNAs. Upon hybridization of the miRNA/messenger (m)RNA duplex, the target mRNA is cleaved and degraded or blocked from translation. The miRNAs represent key players in the regulation of multiple genes, and thus, virtually all cell processes, including cell cycle regulation, apoptosis, differentiation, and metabolism.

MicroRNA sponges are exogenously introduced transcripts containing multiple tandem high-affinity binding sites to a microRNA of interest. These microRNA-binding molecules are competitive regulators that sequester specific microRNAs, thus preventing miRNA/mRNA interaction. MicroRNA sponges are used in loss-of-function studies, where potent miRNA inhibition is desired. It should be noted that microRNA sponges may exhibit off-target effects by sequestering unspecific miRNAs. As the human genome contains thousands of miRNA sequences and given the small size of each mature miRNA (∼22 nucleotides), there is a high probability that poorly designed miRNA sponge constructs may exhibit unspecific sequestration of miRNAs. Therefore, sponge sequences must be carefully optimized to avoid the binding of other miRNAs.

(A) In the absence of sponge treatment, target mRNAs (gray) for a particular miRNA seed family (red complexes) are repressed. Dashed lines indicate mRNA decapping and degradation. (B) After the introduction of the sponge transgene, sponge mRNAs (green) are expressed at a high level and sequester the miRNA complexes, rescuing the expression of the endogenous targets. Sponge-treated cells can be identified by their eGFP reporter expression. (C) The pairing of a miRNA with a bulged sponge site shows mismatches opposite miRNA nucleotides 9–12. The miRNA seed region is highlighted.

Superiority

1. MicroRNA sponge is encoded by a plasmid, which can be easily purified from E.Coli instead of the cost and complex chemical synthesis;
2. MicroRNA sponge is particularly valuable for long-term loss-of-function study both in vitro and in vivo;
3. MicroRNA sponge expression cassette can be engineered into genome by the generation of stable cell lines;
4. Through deliberately designing of microRNA sponge, a whole family of miRNAs can be silenced;
5. Even completely unrelated miRNAs binding sequences can be constructed compatibly within one vector, so microRNA sponge could silence multiple miRNAs simultaneously;
6. Multiple options for miRNA sponge vectors: non-viral/viral plasmid backbone, promoter, reporter gene, antibiotic selection marker, tandem repeat number, etc.;
7. Cost-effective and fast turnaround time;

For more information about our MicroRNA Sponge Service, please call us at 1-862-686-2696 or send an email to [email protected]. Your questions and requests will be answered by expert staff in AcceGen within 24 hours.

MicroRNA Sponge Service

Superiority

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