Functional microRNA screening using a comprehensive lentiviral human microRNA expression library

BMC Genomics

Methodology Article

Begoña Diosdado-Calvo¹  Gerrit A Meijer¹  Lisette M Timmer¹  Anne S Bolijn¹  Jos B Poell²  Eugene Berezikov³  Edwin Cuppen³  Andre AFL van Puijenbroek⁴  Roel QJ Schaapveld⁴  Francesco Cerisoli⁴  Rick J van Haastert⁴ 

[1] Department of Pathology, VU University Medical Center, Amsterdam, The Netherlands;Hubrecht Institute, Cancer Genomics Center, and University Medical Center Utrecht, Uppsalalaan 8, 3584, Utrecht, CT, The Netherlands;Hubrecht Institute, Cancer Genomics Center, and University Medical Center Utrecht, Uppsalalaan 8, 3584, Utrecht, CT, The Netherlands;InteRNA Technologies BV, Padualaan 8, 3585, Utrecht, CH, The Netherlands;InteRNA Technologies BV, Padualaan 8, 3585, Utrecht, CH, The Netherlands;

关键词: HCT15 Cell;    Transduction Efficiency;    Human miRNAs;    Virus Supernatant;    Candidate miRNAs;   

DOI  :  10.1186/1471-2164-12-546

 received in 2011-08-18, accepted in 2011-11-03,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundMicroRNAs (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.ResultsTo 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.ConclusionsWith the miRNA expression library we provide a versatile tool for the systematic elucidation of miRNA function.

【 授权许可】

Unknown   
© Poell et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.