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Simultaneous In Situ Detection of mA-Modified and Unmodified RNAs Using DART-FISH. | LitMetric
N-methyladenosine (mA) is an abundant mRNA modification which plays important roles in regulating RNA function and gene expression. Traditional methods for visualizing mRNAs within cells cannot distinguish mA-modified and unmodified versions of the target transcript, thus limiting our understanding of how and where methylated transcripts are localized within cells. Here, we describe DART-FISH, a visualization technique which enables simultaneous detection of both mA-modified and unmodified target transcripts. DART-FISH combines mA-dependent C-to-U editing with mutation-selective fluorescence in situ hybridization to specifically detect methylated and unmethylated transcript copies, enabling the investigation of mA stoichiometry and methylated mRNA localization in single cells.
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