Abstract
We have recently developed a biochemical approach to isolate miRNA-bound mRNAs and have used this method to identify the genome-wide mRNAs regulated by the tumor suppressor miRNA miR-34a. This method involves transfection of cells with biotinylated miRNA mimics, streptavidin pulldown, RNA isolation, and qRT-PCR. The protocol in this chapter describes these steps and the issues that should be considered while designing such pulldown experiments.
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Acknowledgements
This work was supported by the Intramural Research Program of the National Institutes of Health, National Cancer Institute, Center for Cancer Research.
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Subramanian, M., Li, X.L., Hara, T., Lal, A. (2015). A Biochemical Approach to Identify Direct MicroRNA Targets. In: Carmichael, G. (eds) Regulatory Non-Coding RNAs. Methods in Molecular Biology, vol 1206. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1369-5_3
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DOI: https://doi.org/10.1007/978-1-4939-1369-5_3
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