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Single-molecule fluorescence in situ hybridization: Quantitative imaging of single RNA molecules

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Author(s): Sunjong Kwon

Journal: BMB Reports
ISSN 1976-6696

Volume: 46;
Issue: 2;
Start page: 65;
Date: 2013;
Original page

Keywords: Breast cancer | ERBB2 | FISH | Ribonucleoprotein (RNP) | RNA | Single-molecule imaging

ABSTRACT
In situ detection of RNAs is becoming increasingly importantfor analysis of gene expression within and between intact cellsin tissues. International genomics efforts are now catalogingpatterns of RNA transcription that play roles in cell function,differentiation, and disease formation, and they are demonstratingthe importance of coding and noncoding RNAtranscripts in these processes. However, these techniquestypically provide ensemble averages of transcription acrossmany cells. In situ hybridization-based analysis methodscomplement these studies by providing information about howexpression levels change between cells within normal anddiseased tissues, and they provide information about thelocalization of transcripts within cells, which is important inunderstanding mechanisms of gene regulation. Multi-color,single-molecule fluorescence in situ hybridization (smFISH) isparticularly useful since it enables analysis of several differenttranscripts simultaneously. Combining smFISH with immunofluorescentprotein detection provides additional informationabout the association between transcription level, cellularlocalization, and protein expression in individual cells. [BMBReports 2013; 46(2): 65-72]
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