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dCas12a:Pre-crRNA: A New Tool to Induce mRNA Degradation in Saccharomyces cerevisiae Synthetic Gene Circuits

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Synthetic Biology

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2760))

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Abstract

We describe a new way to trigger mRNA degradation in Saccharomyces cerevisiae synthetic gene circuits. Our method demands to modify either the 5′- or the 3′-UTR that flanks a target gene with elements from the pre-crRNA of type V Cas12a proteins and expresses a DNase-deficient Cas12a (dCas12a). dCas12a recognizes and cleaves the pre-crRNA motifs on mRNA sequences. Our tool does not require complex engineering operations and permits an efficient control of protein expression via mRNA degradation.

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Acknowledgments

We are grateful to the students of the Synthetic Biology lab for their help. We want to thank Xiangyang Zhang and Zhi Li for their assistance in the FACS experiments.

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Authors and Affiliations

  1. School of Pharmaceutical Science and Technology, Tianjin University, Tianjin, China

    Lifang Yu & Mario Andrea Marchisio

Authors
  1. Lifang Yu
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  2. Mario Andrea Marchisio
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Corresponding author

Correspondence to Mario Andrea Marchisio .

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Editors and Affiliations

  1. Agilent Technologies, Inc, La Jolla, CA, USA

    Jeffrey Carl Braman

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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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Yu, L., Marchisio, M.A. (2024). dCas12a:Pre-crRNA: A New Tool to Induce mRNA Degradation in Saccharomyces cerevisiae Synthetic Gene Circuits. In: Braman, J.C. (eds) Synthetic Biology. Methods in Molecular Biology, vol 2760. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3658-9_6

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  • DOI: https://doi.org/10.1007/978-1-0716-3658-9_6

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3657-2

  • Online ISBN: 978-1-0716-3658-9

  • eBook Packages: Springer Protocols

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