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Detection of MicroRNAs Using Synthetic Toehold Switch in Mammalian Cells

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Mammalian Synthetic Systems

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2774))

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Abstract

Engineering synthetic gene circuits to control cellular functions has a broad application in the field of synthetic biology. Synthetic RNA-based switches that can operate at the transcriptional and posttranscriptional level have also drawn significant interest for the application of next-generation therapeutics and diagnostics. Thus, RNA-based switchable platforms are needed to report dynamic cellular mechanisms which play an important role in cell development and diseases. Recently, several RNA-based switches have been designed and utilized for biosensing and molecular diagnostics. However, miRNA-based switches have not been well established or characterized, especially for eukaryotic translational control. Here, we designed a novel synthetic toehold switch for detection of exogenously and endogenously expressed miRNAs in CHO, HeLa, HEK 293, and MDA-MB-231 breast cancer cells. Multiplex detection of miR-155 and miR-21 was tested using two toehold switches to evaluate the orthogonality and programmability of this synthetic platform.

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Author information

Authors and Affiliations

  1. Department of Chemistry, Chemical and Biomedical Engineering, Tagliatela College of Engineering, University of New Haven, West Haven, CT, USA

    Yuwen Zhao, Pratima Poudel & Shue Wang

  2. Department of Bioengineering, Lehigh University, Bethlehem, PA, USA

    Yuwen Zhao

Authors
  1. Yuwen Zhao
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  2. Pratima Poudel
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  3. Shue Wang
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Corresponding author

Correspondence to Shue Wang .

Editor information

Editors and Affiliations

  1. Department of Chemical Engineering and Imperial College Centre for Synthetic Biology, Imperial College London, London, UK

    Francesca Ceroni

  2. Department of Chemical Engineering and Imperial College Centre for Synthetic Biology, Imperial College London, London, UK

    Karen Polizzi

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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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Zhao, Y., Poudel, P., Wang, S. (2024). Detection of MicroRNAs Using Synthetic Toehold Switch in Mammalian Cells. In: Ceroni, F., Polizzi, K. (eds) Mammalian Synthetic Systems. Methods in Molecular Biology, vol 2774. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3718-0_16

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  • DOI: https://doi.org/10.1007/978-1-0716-3718-0_16

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3717-3

  • Online ISBN: 978-1-0716-3718-0

  • eBook Packages: Springer Protocols

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