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Methods Enzymol. 2005;392:371-80.

Use of RNA polymerase II to transcribe artificial microRNAs.

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Department of Molecular Genetics and Microbiology, Howard Hughes Medical Institute, Duke University Medical Center, Durham, NC 27710, USA.


MicroRNAs (miRNAs) are endogenously encoded approximately 22-nt-long RNAs that are generally expressed in a highly tissue- or developmental-stage-specific fashion and that posttranscriptionally regulate target genes. Regulatable RNA polymerase II promoters can be used to overexpress authentic microRNAs in cell culture. Furthermore, one can also design and express artificial microRNAs based on the features of existing microRNA genes, such as the gene encoding the human miR-30 microRNA. Overexpression or inappropriate expression of authentic microRNAs may facilitate the study of their normal functions and expression of artificial microRNAs may permit effective, regulated RNA interference in vivo.

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