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Single-Cell Imaging of mRNA by Target RNA-Initiated RCA

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RNA Amplification and Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2822))

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Abstract

We present a powerful method for direct mRNA detection based on ligation-based recognition and in situ amplification, capable of single-cell imaging mRNA at single-nucleotide and single-molecule resolution. Attributed to the use of Splint R ligase that can ligate padlock probe with RNA as target template, this method can efficiently detect mRNA in the absence of reverse transcription. This method enables spatial localization and correlation analysis of gene expression in single cells, which helps us to elucidate gene function and regulatory mechanisms.

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Acknowledgments

This work was supported by grants from the National Key Research and Development Program of China (2022YFA1603701), National Natural Science Foundation of China (22274004, 22027807), R&D Program of Beijing Municipal Education Commission (KM202310005015), and the Strategic Priority Research Program of Chinese Academy of Sciences (No. XDB36000000).

Author information

Authors and Affiliations

  1. Department of Chemistry, College of Chemistry and Life Science, Beijing University of Technology, Beijing, China

    Xiaojun Ren & Yifan Wu

  2. College of Biomass Science and Engineering, Healthy Food Evaluation Research Center, Sichuan University, Chengdu, China

    Ruijie Deng

  3. Beijing Life Science Academy, Beijing, China

    Jinghong Li

  4. Department of Chemistry, Center for BioAnalytical Chemistry, Key Laboratory of Bioorganic Phosphorus Chemistry & Chemical Biology, Tsinghua University, Beijing, China

    Jinghong Li

  5. New Cornerstone Science Laboratory, Shenzhen, China

    Jinghong Li

  6. Center for BioAnalytical Chemistry, Hefei National Laboratory of Physical Science at Microscale, University of Science and Technology of China, Hefei, China

    Jinghong Li

Authors
  1. Xiaojun Ren
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  2. Yifan Wu
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  3. Ruijie Deng
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  4. Jinghong Li
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Corresponding author

Correspondence to Jinghong Li .

Editor information

Editors and Affiliations

  1. Applied Biology Section, Johns Hopkins University Applied Physic, Laurel, MD, USA

    Mekbib Astatke

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Ren, X., Wu, Y., Deng, R., Li, J. (2024). Single-Cell Imaging of mRNA by Target RNA-Initiated RCA. In: Astatke, M. (eds) RNA Amplification and Analysis. Methods in Molecular Biology, vol 2822. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3918-4_6

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  • DOI: https://doi.org/10.1007/978-1-0716-3918-4_6

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3917-7

  • Online ISBN: 978-1-0716-3918-4

  • eBook Packages: Springer Protocols

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