CRISPR/Cas9-Mediated Gene Editing of the Jasmonate Biosynthesis OsAOC Gene in Rice

  • Trang Hieu Nguyen
  • Huong To Thi Mai
  • Daniel Moukouanga
  • Michel Lebrun
  • Stephane Bellafiore
  • Antony ChampionEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2085)


The function of Jasmonate (JA) is well documented in different plant physiological processes as well as in the interactions with their environment. Mutants impaired in JA production and/or signaling are useful materials to study the function of this phytohormone. Genes involved in the JA biosynthesis pathway in rice have been described, but few mutants deficient in JA production and signaling have been identified. Moreover, these mutants are mostly generated through random mutagenesis approaches, such as irradiation, EMS treatment, or T-DNA insertion, and potentially harbor undesired mutations that could affect other biological processes. The CRISPR/Cas9 system is a precise and efficient genome editing tool that creates DNA modification at specific loci and limit undesired mutations.

In this chapter, we describe a procedure to generate new JA-deficient mutant using CRISPR/Cas9 system in rice. The Allene Oxide Cyclase (OsAOC) gene is targeted since it is a single copy gene in the JA biosynthesis pathway in rice. The widely used variety Oryza sativa japonica Kitaake has been chosen due to its short life cycle and its ease of genetic transformation. This protocol describes the selection of the 20-nt target sequence, construction of the binary vector, and strategy for selecting the T-DNA-free mutant.

Key words

Jasmonate Mutant Rice CRISPR/Cas9 

Supplementary material

464998_2_En_15_MOESM1_ESM.docx (13 kb)
Data 1 (DOCX 13 kb)


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Copyright information

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

Authors and Affiliations

  • Trang Hieu Nguyen
    • 1
  • Huong To Thi Mai
    • 2
  • Daniel Moukouanga
    • 1
  • Michel Lebrun
    • 1
    • 2
    • 3
  • Stephane Bellafiore
    • 4
  • Antony Champion
    • 5
    Email author
  1. 1.Institut de Recherche pour le Developpement (IRD), CiradUniversite fe Montpellier, DIADEMontpellierFrance
  2. 2.Vietnam Academy of Science and Technology (VAST), LMI-RICE2University of Science and Technology of Hanoi (USTH)HanoiVietnam
  3. 3.IRD, CiradUniv Montpellier, LSTMMontpellierFrance
  4. 4.IRD, CiradUniv Montpellier, IPMEMontpellierFrance
  5. 5.UMR DIADE - IRDMontpellierFrance

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