title: Selective gene dosage by CRISPR-Cas9 genome editing in hexaploid Camelina sativa.
abstract: In many plant species, gene dosage is an important cause of phenotype variation. Engineering gene dosage, particularly in polyploid genomes, would provide an efficient tool for plant breeding. The hexaploid oilseed crop Camelina sativa, which has three closely related expressed subgenomes, is an ideal species for investigation of the possibility of creating a large collection of combinatorial mutants. Selective, targeted mutagenesis of the three delta-12-desaturase (FAD2) genes was achieved by CRISPR-Cas9 gene editing, leading to reduced levels of polyunsaturated fatty acids and increased accumulation of oleic acid in the oil. Analysis of mutations over four generations demonstrated the presence of a large variety of heritable mutations in the three isologous CsFAD2 genes. The different combinations of single, double and triple mutants in the T3 generation were isolated, and the complete loss-of-function mutants revealed the importance of delta-12-desaturation for Camelina development. Combinatorial association of different alleles for the three FAD2 loci provided a large diversity of Camelina lines with various lipid profiles, ranging from 10% to 62% oleic acid accumulation in the oil. The different allelic combinations allowed an unbiased analysis of gene dosage and function in this hexaploid species, but also provided a unique source of genetic variability for plant breeding.
authors.affiliation: Institut Jean-Pierre Bourgin (IJPB), INRA, AgroParisTech, CNRS, Saclay Plant Sciences (SPS), Université Paris-Saclay, Versailles, France.
authors.forename: Céline
authors.initials: C
authors.lastname: Morineau
authors.affiliation: Institut Jean-Pierre Bourgin (IJPB), INRA, AgroParisTech, CNRS, Saclay Plant Sciences (SPS), Université Paris-Saclay, Versailles, France.
authors.forename: Yannick
authors.initials: Y
authors.lastname: Bellec
authors.affiliation: Institut Jean-Pierre Bourgin (IJPB), INRA, AgroParisTech, CNRS, Saclay Plant Sciences (SPS), Université Paris-Saclay, Versailles, France.
authors.forename: Frédérique
authors.initials: F
authors.lastname: Tellier
authors.affiliation: Institut Jean-Pierre Bourgin (IJPB), INRA, AgroParisTech, CNRS, Saclay Plant Sciences (SPS), Université Paris-Saclay, Versailles, France.
authors.forename: Lionel
authors.initials: L
authors.lastname: Gissot
authors.affiliation: Institut Jean-Pierre Bourgin (IJPB), INRA, AgroParisTech, CNRS, Saclay Plant Sciences (SPS), Université Paris-Saclay, Versailles, France.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Kelemen
authors.affiliation: Institut Jean-Pierre Bourgin (IJPB), INRA, AgroParisTech, CNRS, Saclay Plant Sciences (SPS), Université Paris-Saclay, Versailles, France.
authors.forename: Fabien
authors.initials: F
authors.lastname: Nogué
authors.affiliation: Institut Jean-Pierre Bourgin (IJPB), INRA, AgroParisTech, CNRS, Saclay Plant Sciences (SPS), Université Paris-Saclay, Versailles, France.
authors.forename: Jean-Denis
authors.initials: JD
authors.lastname: Faure
mesh_terms: D019607:Brassicaceae / Q000235:genetics* / Q000378:metabolism; D064113:CRISPR-Cas Systems / Q000235:genetics*; D044943:Fatty Acid Desaturases / Q000235:genetics / Q000378:metabolism; D018628:Gene Dosage / Q000235:genetics*; D000072669:Gene Editing; D019301:Oleic Acid / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000378:metabolism
chemical_list: D010940:Plant Proteins; D019301:Oleic Acid; D044943:Fatty Acid Desaturases
keywords: CRISPR-Cas9; Camelina; FAD2; oleic acid