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CRISPR/Cas-Mediated In Planta Gene Targeting

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Plant Genomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1610))

Abstract

The recent emergence of the CRISPR/Cas system has boosted the possibilities for precise genome engineering approaches throughout all kingdoms of life. The most common application for plants is targeted mutagenesis, whereby a Cas9-mediated DNA double-strand break (DSB) is repaired by mutagenic nonhomologous end joining (NHEJ). However, the site-specific alteration of a genomic sequence or integration of a transgene relies on the precise repair by homologous recombination (HR) using a suitable donor sequence: this poses a particular challenge in plants, as NHEJ is the preferred repair mechanism for DSBs in somatic tissue. Here, we describe our recently developed in planta gene targeting (ipGT) system, which works via the induction of DSBs by Cas9 to activate the target and the targeting vector at the same time, making it independent of high transformation efficiencies.

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Acknowledgment

We thank Amy Whitbread for the critical reading of the manuscript. Our work on Cas9-mediated genome engineering and GT was funded by the European Research Council (Advanced Grant “COMREC”) as well as the Federal Ministry of Education and Research (PLANT 2030, Pflanzenbiotechnologie fur die Zukunft – TAMOCRO, Grant 0315948).

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Authors and Affiliations

  1. Botanical Institute II, Karlsruhe Institute of Technology, 6980, 76049, Karlsruhe, Germany

    Simon Schiml, Friedrich Fauser & Holger Puchta

  2. Department of Plant Biology, Carnegie Institution for Science, 260 Panama St, Stanford, CA, 94305, USA

    Friedrich Fauser

Authors
  1. Simon Schiml
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  2. Friedrich Fauser
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  3. Holger Puchta
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Corresponding author

Correspondence to Holger Puchta .

Editor information

Editors and Affiliations

  1. Gregor Mendel Institute (GMI), Austrian Academy of Sciences, Vienna Biocenter (VBC), Vienna, Austria

    Wolfgang Busch

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Schiml, S., Fauser, F., Puchta, H. (2017). CRISPR/Cas-Mediated In Planta Gene Targeting. In: Busch, W. (eds) Plant Genomics. Methods in Molecular Biology, vol 1610. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7003-2_1

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  • DOI: https://doi.org/10.1007/978-1-4939-7003-2_1

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7001-8

  • Online ISBN: 978-1-4939-7003-2

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