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Use of the Cas9 Orthologs from Streptococcus thermophilus and Staphylococcus aureus for Non-Homologous End-Joining Mediated Site-Specific Mutagenesis in Arabidopsis thaliana

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Plant Germline Development

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1669))

Abstract

Since the discovery of the CRISPR/Cas system and its in vivo application for site-specific targeted mutagenesis, this technique is wildly used in a great variety of organisms, such as many plant species. Commonly used for this application is the Cas9 enzyme from Streptococcus pyogenes. Here, we describe the application of two Cas9 orthologs from Streptococcus thermophilus and Staphylococcus aureus for targeted non-homologous end-joining mediated mutagenesis in Arabidopsis thaliana. With both orthologs, we could show efficient inheritance of the induced mutations. As both Cas9 orthologs are smaller in size than the enzyme of S. pyogenes and as the Protospacer adjacent motifs (PAMs) differ between all orthologs, they are attractive alternative tools for genome engineering in plants.

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Acknowledgement

The work on our RNA-guided Cas9 vectors was funded by the European Research Council (Advanced Grant “COMREC”).

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Authors and Affiliations

  1. Botanical Institute, Molecular Biology and Biochemistry, Karlsruhe Institute of Technology, 76133, Karlsruhe, Germany

    Jeannette Steinert, Carla Schmidt & Holger Puchta

Authors
  1. Jeannette Steinert
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  2. Carla Schmidt
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  3. Holger Puchta
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Corresponding author

Correspondence to Holger Puchta .

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Editors and Affiliations

  1. Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany

    Anja Schmidt

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Steinert, J., Schmidt, C., Puchta, H. (2017). Use of the Cas9 Orthologs from Streptococcus thermophilus and Staphylococcus aureus for Non-Homologous End-Joining Mediated Site-Specific Mutagenesis in Arabidopsis thaliana . In: Schmidt, A. (eds) Plant Germline Development. Methods in Molecular Biology, vol 1669. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7286-9_27

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  • DOI: https://doi.org/10.1007/978-1-4939-7286-9_27

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7285-2

  • Online ISBN: 978-1-4939-7286-9

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