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A Cas3-base editing tool for targetable in vivo mutagenesis

Anna Zimmermann, Julian E. Prieto-Vivas, Charlotte Cautereels, Anton Gorkovskiy, Jan Steensels, Yves Van de Peer (UGent) and Kevin J. Verstrepen
(2023) NATURE COMMUNICATIONS. 14(1).
Author
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Abstract
The generation of genetic diversity via mutagenesis is routinely used for protein engineering and pathway optimization. Current technologies for random mutagenesis often target either the whole genome or relatively narrow windows. To bridge this gap, we developed CoMuTER (Confined Mutagenesis using a Type I-E CRISPR-Cas system), a tool that allows inducible and targetable, in vivo mutagenesis of genomic loci of up to 55 kilobases. CoMuTER employs the targetable helicase Cas3, signature enzyme of the class 1 type I-E CRISPR-Cas system, fused to a cytidine deaminase to unwind and mutate large stretches of DNA at once, including complete metabolic pathways. The tool increases the number of mutations in the target region 350-fold compared to the rest of the genome, with an average of 0.3 mutations per kilobase. We demonstrate the suitability of CoMuTER for pathway optimization by doubling the production of lycopene in Saccharomyces cerevisiae after a single round of mutagenesis.
Keywords
General Physics and Astronomy, General Biochemistry, Genetics and Molecular Biology, General Chemistry, Multidisciplinary, ESCHERICHIA-COLI, YARROWIA-LIPOLYTICA, DIRECTED EVOLUTION, GENE-EXPRESSION, DNA, DIVERSITY, EFFICIENT, BASE, CLASSIFICATION, DEGRADATION

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Citation

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MLA
Zimmermann, Anna, et al. “A Cas3-Base Editing Tool for Targetable in Vivo Mutagenesis.” NATURE COMMUNICATIONS, vol. 14, no. 1, 2023, doi:10.1038/s41467-023-39087-z.
APA
Zimmermann, A., Prieto-Vivas, J. E., Cautereels, C., Gorkovskiy, A., Steensels, J., Van de Peer, Y., & Verstrepen, K. J. (2023). A Cas3-base editing tool for targetable in vivo mutagenesis. NATURE COMMUNICATIONS, 14(1). https://doi.org/10.1038/s41467-023-39087-z
Chicago author-date
Zimmermann, Anna, Julian E. Prieto-Vivas, Charlotte Cautereels, Anton Gorkovskiy, Jan Steensels, Yves Van de Peer, and Kevin J. Verstrepen. 2023. “A Cas3-Base Editing Tool for Targetable in Vivo Mutagenesis.” NATURE COMMUNICATIONS 14 (1). https://doi.org/10.1038/s41467-023-39087-z.
Chicago author-date (all authors)
Zimmermann, Anna, Julian E. Prieto-Vivas, Charlotte Cautereels, Anton Gorkovskiy, Jan Steensels, Yves Van de Peer, and Kevin J. Verstrepen. 2023. “A Cas3-Base Editing Tool for Targetable in Vivo Mutagenesis.” NATURE COMMUNICATIONS 14 (1). doi:10.1038/s41467-023-39087-z.
Vancouver
1.
Zimmermann A, Prieto-Vivas JE, Cautereels C, Gorkovskiy A, Steensels J, Van de Peer Y, et al. A Cas3-base editing tool for targetable in vivo mutagenesis. NATURE COMMUNICATIONS. 2023;14(1).
IEEE
[1]
A. Zimmermann et al., “A Cas3-base editing tool for targetable in vivo mutagenesis,” NATURE COMMUNICATIONS, vol. 14, no. 1, 2023.
@article{01H43K0JN12AHYX89TT5VQRBYD,
  abstract     = {{The generation of genetic diversity via mutagenesis is routinely used for protein engineering and pathway optimization. Current technologies for random mutagenesis often target either the whole genome or relatively narrow windows. To bridge this gap, we developed CoMuTER (Confined Mutagenesis using a Type I-E CRISPR-Cas system), a tool that allows inducible and targetable, in vivo mutagenesis of genomic loci of up to 55 kilobases. CoMuTER employs the targetable helicase Cas3, signature enzyme of the class 1 type I-E CRISPR-Cas system, fused to a cytidine deaminase to unwind and mutate large stretches of DNA at once, including complete metabolic pathways. The tool increases the number of mutations in the target region 350-fold compared to the rest of the genome, with an average of 0.3 mutations per kilobase. We demonstrate the suitability of CoMuTER for pathway optimization by doubling the production of lycopene in Saccharomyces cerevisiae after a single round of mutagenesis.}},
  articleno    = {{3389}},
  author       = {{Zimmermann, Anna and Prieto-Vivas, Julian E. and Cautereels, Charlotte and Gorkovskiy, Anton and Steensels, Jan and Van de Peer, Yves and Verstrepen, Kevin J.}},
  issn         = {{2041-1723}},
  journal      = {{NATURE COMMUNICATIONS}},
  keywords     = {{General Physics and Astronomy,General Biochemistry, Genetics and Molecular Biology,General Chemistry,Multidisciplinary,ESCHERICHIA-COLI,YARROWIA-LIPOLYTICA,DIRECTED EVOLUTION,GENE-EXPRESSION,DNA,DIVERSITY,EFFICIENT,BASE,CLASSIFICATION,DEGRADATION}},
  language     = {{eng}},
  number       = {{1}},
  pages        = {{16}},
  title        = {{A Cas3-base editing tool for targetable in vivo mutagenesis}},
  url          = {{http://doi.org/10.1038/s41467-023-39087-z}},
  volume       = {{14}},
  year         = {{2023}},
}
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