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Open Access Highly Accessed Methodology article

Functional microRNA screening using a comprehensive lentiviral human microRNA expression library

Jos B Poell1, Rick J van Haastert2, Francesco Cerisoli2, Anne S Bolijn3, Lisette M Timmer3, Begoña Diosdado-Calvo3, Gerrit A Meijer3, Andre AFL van Puijenbroek2, Eugene Berezikov12, Roel QJ Schaapveld2 and Edwin Cuppen12*

Author Affiliations

1 Hubrecht Institute, Cancer Genomics Center, and University Medical Center Utrecht. Uppsalalaan 8, 3584 CT Utrecht, The Netherlands

2 InteRNA Technologies BV, Padualaan 8, 3585 CH Utrecht, The Netherlands

3 Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands

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BMC Genomics 2011, 12:546  doi:10.1186/1471-2164-12-546

Published: 3 November 2011

Abstract

Background

MicroRNAs (miRNAs) are a class of small regulatory RNAs that target sequences in messenger RNAs (mRNAs) to inhibit their protein output. Dissecting the complexities of miRNA function continues to prove challenging as miRNAs are predicted to have thousands of targets, and mRNAs can be targeted by dozens of miRNAs.

Results

To systematically address biological function of miRNAs, we constructed and validated a lentiviral miRNA expression library containing 660 currently annotated and 422 candidate human miRNA precursors. The miRNAs are expressed from their native genomic backbone, ensuring physiological processing. The arrayed layout of the library renders it ideal for high-throughput screens, but also allows pooled screening and hit picking. We demonstrate its functionality in both short- and long-term assays, and are able to corroborate previously described results of well-studied miRNAs.

Conclusions

With the miRNA expression library we provide a versatile tool for the systematic elucidation of miRNA function.