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2016 ; 1402
(ä): 147-164
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Visualizing Long Noncoding RNAs on Chromatin
#MMPMID26721489
Hinten M
; Maclary E
; Gayen S
; Harris C
; Kalantry S
Methods Mol Biol
2016[]; 1402
(ä): 147-164
PMID26721489
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Fluorescence in situ hybridization (FISH) enables the detection of specific
nucleic acid sequences within single cells. For example, RNA FISH provides
information on both the expression level and localization of RNA transcripts and,
when combined with detection of associated proteins and chromatin modifications,
can lend essential insights into long noncoding RNA (lncRNA) function. Epigenetic
effects have been postulated for many lncRNAs, but shown for only a few. Advances
in in situ techniques and microscopy, however, now allow for visualization of
lncRNAs that are expressed at very low levels or are not very stable. FISH-based
detections of RNA and DNA coupled with immunological staining of proteins/histone
modifications offer the possibility to connect lncRNAs to epigenetic effects.
Here, we describe an integrated set of protocols to detect, individually or in
combination, specific RNAs, DNAs, proteins, and histone modifications in single
cells at a high level of sensitivity using conventional fluorescence microscopy.
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