Generation of microRNA Sponge Library.

Bio-protocol (2018-04-20)
Sebastian Herzog

This protocol describes the generation and functional validation of microRNA (miRNA) sponge or decoy constructs. When expressed from a strong promoter, these transcripts can sequester specific miRNA:RISC complexes, thereby resulting in a derepression of endogenous target mRNA. Hence, cells expressing such sponges display a partial or full miRNA loss-of-function phenotype. Depending on the sponge sequence, the activity of any miRNA of choice can be inhibited by sponge sequestration, but it should be noted that these constructs do not seem to be specific for one particular miRNA. Rather, all miRNAs of the same family as defined by the seed sequence will be affected, albeit to a different degree.

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Trizma® base, Primary Standard and Buffer, ≥99.9% (titration), crystalline
Ethylenediaminetetraacetic acid disodium salt dihydrate, suitable for electrophoresis, for molecular biology, 99.0-101.0% (titration)