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

  1. PETER T. NELSON1,
  2. DON A. BALDWIN1,
  3. WIGARD P. KLOOSTERMAN2,
  4. SAKARI KAUPPINEN3,
  5. RONALD H.A. PLASTERK2, and
  6. ZISSIMOS MOURELATOS1
  1. 1Division of Neuropathology and Department of Pathology & Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA
  2. 2Hubrecht Laboratory, Centre for Biomedical Genetics, Utrecht, The Netherlands
  3. 3Wilhelm Johannsen Centre for Functional Genome Research, Institute of Medical Biochemistry and Genetics, University of Copenhagen, Denmark

Abstract

microRNAs (miRNAs) are small (~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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  1. Published in Advance December 22, 2005, doi: 10.1261/rna.2258506 RNA 12: 187-191 Copyright 2006 by RNA Society

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