RAKE and LNA-ISH reveal microRNA expression and localization in archival human brain.

RNA

Division of Neuropathology and Department of Pathology & Laboratory Medicine, University of Pennsylvania School of Medicine, 605A Stellar-Chance Bldg., 421 Curie Blvd., Philadelphia, PA 19104, USA.

Published: February 2006

microRNAs (miRNAs) are small (approximately 22 nucleotide) regulatory RNAs which play fundamental roles in many biological processes. Recent studies have shown that the expression of many miRNAs is altered in various human tumors and some miRNAs may function as oncogenes or tumor suppressor genes. However, with the exception of glioblastoma multiforme, the expression of miRNAs in brain tumors is unknown. Furthermore, methods to profile miRNAs from formalin-fixed, paraffin-embedded (FFPE) archival tissues or to study their cellular and subcellular localization in FFPE tissues have been lacking. Here we report the coordinated miRNA expression analysis from the tissue level to the subcellular level, using the RAKE (RNA-primed, array-based, Klenow Enzyme) miRNA microarray platform in conjunction with Locked Nucleic Acid (LNA)-based in situ hybridization (LNA-ISH) on archival FFPE human brains and oligodendroglial tumors. The ability to profile miRNAs from archival tissues at the tissue level, by RAKE microarrays, and at the cellular level by LNA-ISH, will accelerate studies of miRNAs in human diseases.

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1370897PMC
http://dx.doi.org/10.1261/rna.2258506DOI Listing

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