title: Genome engineering empowers the diatom Phaeodactylum tricornutum for biotechnology.
abstract: Diatoms, a major group of photosynthetic microalgae, have a high biotechnological potential that has not been fully exploited because of the paucity of available genetic tools. Here we demonstrate targeted and stable modifications of the genome of the marine diatom Phaeodactylum tricornutum, using both meganucleases and TALE nucleases. When nuclease-encoding constructs are co-transformed with a selectable marker, high frequencies of genome modifications are readily attained with 56 and 27% of the colonies exhibiting targeted mutagenesis or targeted gene insertion, respectively. The generation of an enhanced lipid-producing strain (45-fold increase in triacylglycerol accumulation) through the disruption of the UDP-glucose pyrophosphorylase gene exemplifies the power of genome engineering to harness diatoms for biofuel production.
authors.affiliation: Cellectis S.A., 8 rue de la Croix de Jarry, 75013 Paris, France.
authors.forename: Fayza
authors.initials: F
authors.lastname: Daboussi
authors.affiliation: Cellectis S.A., 8 rue de la Croix de Jarry, 75013 Paris, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Leduc
authors.affiliation: Cellectis S.A., 8 rue de la Croix de Jarry, 75013 Paris, France.
authors.forename: Alan
authors.initials: A
authors.lastname: Maréchal
authors.affiliation: Cellectis S.A., 8 rue de la Croix de Jarry, 75013 Paris, France.
authors.forename: Gwendoline
authors.initials: G
authors.lastname: Dubois
authors.affiliation: Cellectis S.A., 8 rue de la Croix de Jarry, 75013 Paris, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Guyot
authors.affiliation: Cellectis S.A., 8 rue de la Croix de Jarry, 75013 Paris, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Perez-Michaut
authors.affiliation: Cellectis S.A., 8 rue de la Croix de Jarry, 75013 Paris, France.
authors.forename: Alberto
authors.initials: A
authors.lastname: Amato
authors.affiliation: 1] Université Pierre et Marie Curie, UMR7238, CNRS-UPMC, 75006 Paris, France [2] CNRS, UMR7238, Laboratoire Biologie Computationnelle et Quantitative, 75006 Paris, France.
authors.forename: Angela
authors.initials: A
authors.lastname: Falciatore
authors.affiliation: Cellectis S.A., 8 rue de la Croix de Jarry, 75013 Paris, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Juillerat
authors.affiliation: Cellectis S.A., 8 rue de la Croix de Jarry, 75013 Paris, France.
authors.forename: Marine
authors.initials: M
authors.lastname: Beurdeley
authors.affiliation: Cellectis plant sciences, 600 County Rd D Suite 8, New Brighton, Minnesota 55112, USA.
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Voytas
authors.affiliation: Cellectis S.A., 8 rue de la Croix de Jarry, 75013 Paris, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Cavarec
authors.affiliation: Cellectis S.A., 8 rue de la Croix de Jarry, 75013 Paris, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Duchateau
mesh_terms: D001483:Base Sequence; D001709:Biotechnology; D002851:Chromatography, High Pressure Liquid; D053903:DNA Breaks, Double-Stranded; D017377:Diatoms / Q000235:genetics*; D004720:Endonucleases / Q000378:metabolism; D005434:Flow Cytometry; D005818:Genetic Engineering / Q000379:methods*; D016678:Genome; D005838:Genotype; D008055:Lipids / Q000096:biosynthesis; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D059645:Mutation Rate; D053719:Tandem Mass Spectrometry; D015534:Trans-Activators / Q000378:metabolism; D014280:Triglycerides / Q000032:analysis
chemical_list: D008055:Lipids; D015534:Trans-Activators; D014280:Triglycerides; D004720:Endonucleases