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MicroRNA profiling of the murine hematopoietic system

DOI: 10.1186/gb-2005-6-8-r71

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Abstract:

We used oligonucleotide arrays to analyze miRNA expression in the murine hematopoietic system. Complementary oligonucleotides capable of hybridizing to 181 miRNAs were immobilized on a membrane and probed with radiolabeled RNA derived from low molecular weight fractions of total RNA from several different hematopoietic and neuronal cells. This method allowed us to analyze cell type-specific patterns of miRNA expression and to identify miRNAs that might be important for cell lineage specification and/or cell effector functions.This is the first report of systematic miRNA gene profiling in cells of the hematopoietic system. As expected, miRNA expression patterns were very different between hematopoietic and non-hematopoietic cells, with further subtle differences observed within the hematopoietic group. Interestingly, the most pronounced similarities were observed among fully differentiated effector cells (Th1 and Th2 lymphocytes and mast cells) and precursors at comparable stages of differentiation (double negative thymocytes and pro-B cells), suggesting that in addition to regulating the process of commitment to particular cellular lineages, miRNAs might have an important general role in the mechanism of cell differentiation and maintenance of cell identity.MicroRNAs (miRNAs) represent a recently discovered class of small, noncoding RNAs, found in organisms ranging from nematodes to plants to humans. Many individual miRNAs are conserved across widely diverse phyla, indicating their physiological importance. The primary transcript (pri-miRNA) is generally transcribed by RNA polymerase II; it contains a typical stem-loop structure that is processed by a nuclear enzyme complex including Drosha and Pasha, and releases a 60- to 110-nucleotide pre-miRNA hairpin precursor [1]. The pre-miRNA is further processed by Dicer to yield the 19- to 22-nucleotide mature miRNA product, which is then incorporated into the RNA-induced silencing complex (RISC) [2-4]. RISC-bound miRNAs d

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