Nguyen T. H., To Thi Mai H., Moukouanga Daniel, Lebrun Michel, Bellafiore Stéphane, Champion Antony. (2020). CRISPR/Cas9-mediated gene editing of the jasmonate biosynthesis OsAOC gene in rice. In :
Champion Antony (ed.), Laplaze Laurent (ed.). Jasmonate in plant biology : methods and protocols. New York : Humana Press, 199-209. (Methods in Molecular Biology). ISBN 978-1-0716-0141-9.
Titre du document
CRISPR/Cas9-mediated gene editing of the jasmonate biosynthesis OsAOC gene in rice
Nguyen T. H., To Thi Mai H., Moukouanga Daniel, Lebrun Michel, Bellafiore Stéphane, Champion Antony
In
Champion Antony (ed.), Laplaze Laurent (ed.), Jasmonate in plant biology : methods and protocols
Source
New York : Humana Press, 2020,
199-209 (Methods in Molecular Biology). ISBN 978-1-0716-0141-9
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 ricehave 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 toits short life cycle and its ease of genetic transformation. This protocol describes the selection of the 20-nttarget sequence, construction of the binary vector, and strategy for selecting the T-DNA-free mutant.
Plan de classement
Sciences du monde végétal [076]
;
Biotechnologies [084]
