Abstract
Long non-coding RNAs (lncRNAs) have been postulated to function in a number of DNA-based processes, most notably transcription. The detection of lncRNAs in situ can offer insights into their function. Fluorescence in situ hybridization (FISH) enables the detection of specific nucleic acid sequences, including lncRNAs, within individual cells. Current RNA FISH techniques can inform both the localization and expression level of RNA transcripts. Together with advances in microscopy, these in situ techniques now allow for visualization and quantification of even lowly expressed or unstable lncRNAs. When combined with detection of associated proteins and chromatin modifications by immunofluorescence, RNA FISH can lend essential insights into lncRNA function. 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 high sensitivity using conventional fluorescence microscopy.
Megan Trotter, Clair Harris, Marissa Cloutier, and Milan Samanta are equal authors.
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Acknowledgments
This work was funded by NIH National Research Service Awards 5-T32-GM07544 (University of Michigan Predoctoral Genetics Training Program to M.C. and M.T.); a T32-HD079342 (University of Michigan Predoctoral Career Training in the Reproductive Sciences Program to M.C.); an NIH Director’s New Innovator award (DP2-OD-008646-01), an NIH NIGMS R01 award (R01GM124571), and an NIH NICHD R01 award (R01HD095463) (to S.K.); and the University of Michigan Endowment for Basic Sciences (to S.K.).
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© 2021 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
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Trotter, M., Harris, C., Cloutier, M., Samanta, M., Kalantry, S. (2021). Highly Resolved Detection of Long Non-coding RNAs In Situ. In: Zhang, L., Hu, X. (eds) Long Non-Coding RNAs. Methods in Molecular Biology, vol 2372. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1697-0_12
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DOI: https://doi.org/10.1007/978-1-0716-1697-0_12
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Online ISBN: 978-1-0716-1697-0
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