Single microRNA (miRNA) can be inhibited using antagomiR which efficiently knockdown a specific miRNA. However, the effect is transient and often results in subtle phenotype. Here we report a guideline on designing miRNA sponge inhibiting a miRNA family. As a model system, we targeted miR-30 family, known as tumor suppressor miRNAs in multiple tumors. To achieve an efficient knockdown, we generated perfect and bulged-matched miRNA binding sites (MBS) and introduced multiple copies of MBS. The protocol here demonstrates the miRNA sponge as a useful tool to examine the functional impact of inhibition miRNAs.
Ortega, M. M., & Bouamar, H. (2017). Guidelines on designing microRNA sponges: From construction to stable cell line. In Methods in Molecular Biology (Vol. 1509, pp. 221–233). Humana Press Inc. https://doi.org/10.1007/978-1-4939-6524-3_20