Robust one-day in situ hybridization protocol for detection of microRNAs in paraffin samples using LNA probes

Methods : a Companion to Methods in Enzymology
Stine JørgensenBoye Schnack Nielsen

Abstract

MicroRNAs (miRNAs) constitute a group of small non-coding RNA molecules generally 18-22 base-pairs in length. miRNAs are considered to be negative regulators of gene expression at the level of post-transcription and are important in normal physiological development, tissue homeostasis and disease. The cellular origin of individual microRNAs is of utmost importance for understanding their roles in molecular and biological processes, in multi-cellular and complex structured tissues. For the localization of miRNAs in clinical and experimental formalin-fixed and paraffin-embedded samples we have developed a simple and robust one-day in situ hybridization protocol based on the use of double digoxigenin (DIG)-labeled LNA-DNA chimeric probes. We show that the protocol enables analyses of specificity, and demonstrate the detection of miR-1, miR-21, miR-124, miR-126, miR-145, and miR-205 in human and murine paraffin material. The well established localization of these microRNAs makes them ideal for use as reference microRNAs when optimizing the microRNA in situ hybridization assay for a particular tissue and miRNA.

References

Jan 17, 2003·Expert Review of Molecular Diagnostics·Peter MouritzenSøren Møller
May 4, 2006·Nucleic Acids Research·Yong YouRichard Owczarzy
Sep 26, 2007·International Journal of Cancer. Journal International Du Cancer·S Perwez Hussain, Curtis C Harris
Jun 12, 2009·Nucleic Acids Research·Jing Lu, Andrew Tsourkas

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Citations

Dec 11, 2013·ACS Chemical Biology·Ponnusamy Babu
Nov 22, 2013·Journal of Experimental Botany·Reidunn Birgitta Aalen
May 6, 2011·Nanomedicine·Mithun M ShenoiJohn C Bischof
Mar 1, 2014·BMC Cancer·Helge StenvoldRoy M Bremnes
Apr 27, 2012·Cellular and Molecular Life Sciences : CMLS·Anne A KanR Jeroen Pasterkamp
Dec 5, 2014·Future Oncology·Marta UsóCarlos Camps
Nov 12, 2010·Clinical & Experimental Metastasis·Boye Schnack NielsenHans Jørgen Nielsen
Apr 29, 2011·The Journal of Histochemistry and Cytochemistry : Official Journal of the Histochemistry Society·Martin J SøeKim Holmstrøm
Nov 4, 2011·BMC Medical Genomics·Matthew N McCallMarc K Halushka
Dec 20, 2011·Nature Cell Biology·A BroniszM C Ostrowski
Jun 13, 2015·International Journal of Molecular Sciences·Martyna O UrbanekWlodzimierz J Krzyzosiak
Apr 17, 2013·Connective Tissue Research·Søren Ohrt-NissenBenny Dahl
Feb 6, 2016·Annals of Surgical Oncology·Christina L RolandKeila E Torres
Jan 9, 2016·Modern Pathology : an Official Journal of the United States and Canadian Academy of Pathology, Inc·Gerald SaldanhaJ Howard Pringle
Nov 28, 2013·Translational Research : the Journal of Laboratory and Clinical Medicine·Kiseok JangSe Jin Jang
Jan 23, 2016·Progress in Histochemistry and Cytochemistry·Anthony Warford
Dec 14, 2011·Heart, Lung & Circulation·Bianca C BernardoJulie R McMullen
Mar 8, 2011·Trends in Molecular Medicine·Cathy A AndorferEdith A Perez
Mar 20, 2012·The American Journal of Pathology·Patricia Puerta-GilMarta Sánchez-Carbayo
Sep 16, 2011·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·Lene RaskBoye Schnack Nielsen
Jun 28, 2011·The Journal of Pathology·Inge de KrijgerIris D Nagtegaal
Nov 10, 2012·International Journal of Experimental Pathology·Denise YanXue Z Liu
May 28, 2011·International Journal of Cancer. Journal International Du Cancer·Benjamin BoyerinasMarcus E Peter
Oct 1, 2014·Transplant International : Official Journal of the European Society for Organ Transplantation·Jianguo ChenRoslyn B Mannon
Mar 13, 2014·Colorectal Disease : the Official Journal of the Association of Coloproctology of Great Britain and Ireland·A BhanguR Goldin
Dec 3, 2014·The American Journal of Pathology·Todd A MacKenzieLorenzo F Sempere
May 3, 2015·Journal of Clinical Pathology·Ambra Vittoria GualeniAnnunziata Gloghini
Oct 11, 2013·Biotechnic & Histochemistry : Official Publication of the Biological Stain Commission·U SinghR Garrido

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