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Live-Cell Imaging of Long Noncoding RNAs Using Molecular Beacons

  • Yachen Ying
  • Shiqi Mao
  • Christopher J. Krueger
  • Antony K. ChenEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2038)

Abstract

Long noncoding RNAs (lncRNAs) are a family of non-protein-coding RNA transcripts greater than 200 nucleotides in length that have been regarded as crucial modulators of gene expression in various biological and disease contexts, but mechanisms underlying such regulation still remains largely elusive. In addition to cell lysate-based approaches that have proven invaluable for studies of lncRNAs, live-imaging methods can add value by providing more in-depth information on lncRNA dynamics and localizations at the single-molecule level. Recently, we have developed a versatile imaging approach based on molecular beacons (MBs), which are a class of fluorogenic oligonucleotide-based probes with the capacity to convert RNA target hybridization into a measurable fluorescence signal. In this chapter, we describe the detailed protocol of using MBs to illuminate lncRNA transcripts at the single-molecule level in living cells.

Key words

Molecular beacons lncRNA Tandem repeats Single-molecule 

Notes

Acknowledgments

This project was supported by grants from the National Key R&D Program of China (Nos. 2016YFA0501603 and 2016YFA0100702), the National Natural Science Foundation of China (Nos. 31771583 and 81371613), and the Beijing Natural Science Foundation (7162114).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Yachen Ying
    • 1
  • Shiqi Mao
    • 1
  • Christopher J. Krueger
    • 1
    • 2
  • Antony K. Chen
    • 1
    Email author
  1. 1.Department of Biomedical EngineeringCollege of Engineering, Peking UniversityBeijingChina
  2. 2.Wallace H Coulter Department of Biomedical EngineeringGeorgia Institute of TechnologyAtlantaUSA

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