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Department of Molecular Biomedicine, Institute of Bioorganic Chemistry, Polish Academy of Sciences, Noskowskiego 12-14 Str., 61-704 Poznan, Poland



These authors contributed equally to this work.





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Academic Editor: Michael Ladomery

Abstract Small noncoding RNAs perform multiple regulatory functions in cells, and their exogenous mimics are widely used in research and experimental therapies to interfere with target gene expression. MicroRNAs miRNAs are the most thoroughly investigated representatives of the small RNA family, which includes short interfering RNAs siRNAs, PIWI-associated RNA piRNAs, and others. Numerous methods have been adopted for the detection and characterization of small RNAs, which is challenging due to their short length and low level of expression. These include molecular biology methods such as real-time RT-PCR, northern blotting, hybridization to microarrays, cloning and sequencing, as well as single cell miRNA detection by microscopy with in situ hybridization ISH. In this review, we focus on the ISH method, including its fluorescent version FISH, and we present recent methodological advances that facilitated its successful adaptation for small RNA detection. We discuss relevant technical aspects as well as the advantages and limitations of ISH. We also refer to numerous applications of small RNA ISH in basic research and molecular diagnostics. View Full-Text

Keywords: short interfering RNA; Piwi-interacting RNA; LNA probe; rolling circle amplification; padlock probes; enzyme-labeled fluorescence signal amplification; TIRCA; PLA; tyramide signal amplification short interfering RNA; Piwi-interacting RNA; LNA probe; rolling circle amplification; padlock probes; enzyme-labeled fluorescence signal amplification; TIRCA; PLA; tyramide signal amplification





Autor: Martyna O. Urbanek †, Anna U. Nawrocka † and Wlodzimierz J. Krzyzosiak *

Fuente: http://mdpi.com/



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