Patterns of CRISPR/Cas9 activity in plants, animals and microbes

  • Luisa Bortesi
    Institute for Molecular Biotechnology RWTH Aachen University Aachen Germany
  • Changfu Zhu
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Julia Zischewski
    Institute for Molecular Biotechnology RWTH Aachen University Aachen Germany
  • Lucia Perez
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Ludovic Bassié
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Riad Nadi
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Giobbe Forni
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Sarah Boyd Lade
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Erika Soto
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Xin Jin
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Vicente Medina
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Gemma Villorbina
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Pilar Muñoz
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Gemma Farré
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Rainer Fischer
    Institute for Molecular Biotechnology RWTH Aachen University Aachen Germany
  • Richard M. Twyman
    TRM Ltd York UK
  • Teresa Capell
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Paul Christou
    Department of Plant Production and Forestry Science School of Agrifood and Forestry Science and Engineering (ETSEA) University of Lleida‐Agrotecnio Center Lleida Spain
  • Stefan Schillberg
    Fraunhofer Institute for Molecular Biology and Applied Ecology IME Aachen Germany

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<jats:title>Summary</jats:title><jats:p>The CRISPR/Cas9 system and related RNA‐guided endonucleases can introduce double‐strand breaks (DSBs) at specific sites in the genome, allowing the generation of targeted mutations in one or more genes as well as more complex genomic rearrangements. Modifications of the canonical CRISPR/Cas9 system from <jats:italic>Streptococcus pyogenes</jats:italic> and the introduction of related systems from other bacteria have increased the diversity of genomic sites that can be targeted, providing greater control over the resolution of DSBs, the targeting efficiency (frequency of on‐target mutations), the targeting accuracy (likelihood of off‐target mutations) and the type of mutations that are induced. Although much is now known about the principles of CRISPR/Cas9 genome editing, the likelihood of different outcomes is species‐dependent and there have been few comparative studies looking at the basis of such diversity. Here we critically analyse the activity of CRISPR/Cas9 and related systems in different plant species and compare the outcomes in animals and microbes to draw broad conclusions about the design principles required for effective genome editing in different organisms. These principles will be important for the commercial development of crops, farm animals, animal disease models and novel microbial strains using CRISPR/Cas9 and other genome‐editing tools.</jats:p>

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