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Precursor miRNAs for Successful miRNA Functional Studies

Pre-miR miRNA Precursor Molecules
miRNA target site identification and validation
Screening for miRNAs that regulate target gene expression
Screening for miRNAs that affect a cellular process


MicroRNAs (miRNAs) are small, regulatory RNAs that are expressed in animals and plants and affect the translation or stability of target mRNAs. The 17-24 nt, single-stranded (ss) miRNAs are derived from longer, primary transcripts termed "pri-miRNAs" [1]. The pri-miRNAs, which can be more than 1000 nt in length, contain an RNA hairpin in which one of the two strands includes the mature miRNA [1]. The hairpin, which typically comprises 60-120 nt, is cleaved from the pri-miRNA in the nucleus by the double-strand-specific ribonuclease, Drosha [1]. The resulting precursor miRNA, or "pre-miRNA," is transported to the cytoplasm via a process that involves Exportin-5 [2-4]. The pre-miRNA is further cleaved by Dicer [5] to generate a short, partially double-stranded (ds) RNA in which one strand is the mature miRNA. The mature miRNA is taken up by a protein complex that is similar, if not identical, to the RNA Induced Silencing Complex (RISC) that supports RNA interference (RNAi) [6], and miRNA-bound complex functions to regulate translation.

miRNA functional analyses can be carried out using tools that are similar to those used to analyze protein encoding genes. Up-regulation of the miRNAs
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