A Sensitive Alternative for MicroRNA In Situ Hybridizations Using Probes of 2′-O-Methyl RNA plus LNA

被引：38

作者：

Soe, Martin J. ^{[1
]}

Moller, Trine ^{[1
]}

Dufva, Martin ^{[2
]}

Holmstrom, Kim ^{[1
]}

机构：

[1] Bioneer AS, DK-2970 Horsholm, Denmark

[2] Tech Univ Denmark, DTU Nanotech, DK-2800 Lyngby, Denmark

来源：

JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY | 2011年 / 59卷 / 07期

关键词：

in situ hybridization; microRNA; locked nucleic acids; 2 '-O-methyl RNA; yeast RNA; urea; formalin-fixed paraffin-embedded; microRNA-138; microRNA-205; LOCKED NUCLEIC-ACID; EXPRESSION; PRECURSOR; REVEALS;

D O I：

10.1369/0022155411409411

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

The use of short, high-affinity probes consisting of a combination of DNA and locked nucleic acid (LNA) has enabled the specific detection of microRNAs (miRNAs) by in situ hybridization (ISH). However, detection of low-copy number miRNAs is still not always possible. Here the authors show that probes consisting of 2'-O-methyl RNAs (2OMe) and LNA at every third base (2:1 ratio), under optimized hybridization conditions, excluding yeast RNA from the hybridization buffer, can provide superior performance in detection of miRNA targets in terms of sensitivity and signal-to-noise ratio compared to DNA + LNA probes. Furthermore, they show that hybridizations can be performed in buffers of 4M urea instead of 50% formamide, thereby yielding an equally specific but nontoxic assay. The use of 2OMe + LNA-based probes and the optimized ISH assay enable simple and fast detection of low-copy number miRNA targets, such as miR-130a in mouse brain. (J Histochem Cytochem 59:661-672, 2011)

引用

页码：661 / 672

页数：12

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