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Gene Excision by Dual-Guide CRISPR-Cas9

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Yarrowia lipolytica

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2307))

Abstract

CRISPR-Cas9 is frequently used for creating double-strand DNA breaks that result in indels through non-homologous end joining. Indels can revert to wild-type sequence and require sequencing or complex assays to measure. Cutting by two guide RNAs can lead to single indels at either cut site or simultaneous cutting at both sites and repair leading to gene excision.

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Acknowledgments

This work was supported by the National Science Foundation (NSF CBET 1403099/1706134) and by an Early Career Award from NASA’s Space Technology Research Grants program.

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Author notes

  1. Mark Blenner

    Present address: Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE, USA

Authors and Affiliations

  1. Department of Chemical and Biomolecular Engineering, Clemson University, Clemson, SC, USA

    Michael Spagnuolo & Mark Blenner

Authors
  1. Michael Spagnuolo
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  2. Mark Blenner
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Corresponding author

Correspondence to Mark Blenner .

Editor information

Editors and Affiliations

  1. Chemical and Environmental Engineering, University of California Riverside, Riverside, CA, USA

    Ian Wheeldon

  2. Department of Chemical and Biomolecular Engineering, Clemson University, Clemson, SC, USA

    Mark Blenner

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Spagnuolo, M., Blenner, M. (2021). Gene Excision by Dual-Guide CRISPR-Cas9. In: Wheeldon, I., Blenner, M. (eds) Yarrowia lipolytica. Methods in Molecular Biology, vol 2307. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1414-3_5

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  • DOI: https://doi.org/10.1007/978-1-0716-1414-3_5

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1413-6

  • Online ISBN: 978-1-0716-1414-3

  • eBook Packages: Springer Protocols

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