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CRISPR-Cas-Mediated Gene Knockout in Tomato

  • Gwen Swinnen
  • Thomas Jacobs
  • Laurens Pauwels
  • Alain GoossensEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2083)

Abstract

Loss-of-function mutants are crucial for plant functional genomics studies. With the advent of CRISPR-Cas genome editing, generating null alleles for one or multiple specific gene(s) has become feasible for many plant species including tomato (Solanum lycopersicum). An easily programmable RNA-guided Cas endonuclease efficiently creates DNA double-strand breaks (DSBs) at targeted genomic sites that can be repaired by nonhomologous end joining (NHEJ) typically leading to small insertions or deletions that can produce null mutations. Here, we describe how to utilize CRISPR-Cas genome editing to obtain stable tomato gene knockout lines.

Key words

Genome editing CRISRP-Cas Gene knockout Loss-of-function mutation Null mutation Site-directed mutagenesis Targeted mutagenesis Tomato Solanum lycopersicum Solanaceae 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Gwen Swinnen
    • 1
    • 2
  • Thomas Jacobs
    • 1
    • 2
  • Laurens Pauwels
    • 1
    • 2
  • Alain Goossens
    • 1
    • 2
    Email author
  1. 1.Department of Plant Biotechnology and BioinformaticsGhent UniversityGhentBelgium
  2. 2.VIB Center for Plant Systems BiologyGhentBelgium

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