title: Localized egg-cell expression of effector proteins for targeted modification of the Arabidopsis genome.
abstract: Targeted modification of the genome is an important genetic tool, which can be achieved via homologous, non-homologous or site-specific recombination. Although numerous efforts have been made, such a tool does not exist for routine applications in plants. This work describes a simple and useful method for targeted mutagenesis or gene targeting, tailored to floral-dip transformation in Arabidopsis, by means of specific protein expression in the egg cell. Proteins stably or transiently expressed under the egg apparatus-specific enhancer (EASE) were successfully localized to the area of the egg cell. Moreover, a zinc-finger nuclease expressed under EASE induced targeted mutagenesis. Mutations obtained under EASE control corresponded to genetically independent events that took place specifically in the germline. In addition, RAD54 expression under EASE led to an approximately 10-fold increase in gene targeting efficiency, when compared with wild-type plants. EASE-controlled gene expression provides a method for the precise engineering of the Arabidopsis genome through temporally and spatially controlled protein expression. This system can be implemented as a useful method for basic research in Arabidopsis, as well as in the optimization of tools for targeted genetic modifications in crop plants.
authors.affiliation: Department of Plant Sciences, The Weizmann Institute of Science, Rehovot, Israel.
authors.forename: Liron
authors.initials: L
authors.lastname: Even-Faitelson
authors.forename: Aviva
authors.initials: A
authors.lastname: Samach
authors.forename: Cathy
authors.initials: C
authors.lastname: Melamed-Bessudo
authors.forename: Naomi
authors.initials: N
authors.lastname: Avivi-Ragolsky
authors.forename: Avraham A
authors.initials: AA
authors.lastname: Levy
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D004265:DNA Helicases / Q000235:genetics / Q000378:metabolism; D003851:Deoxyribonucleases / Q000235:genetics / Q000378:metabolism; D004789:Enzyme Activation; D018506:Gene Expression Regulation, Plant; D018390:Gene Targeting / Q000379:methods*; D017343:Genes, Plant; D005818:Genetic Engineering / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D018745:Genome, Plant; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000379:methods; D056771:Ovule / Q000166:cytology / Q000235:genetics* / Q000378:metabolism; D059828:Plant Cells / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D015533:Transcriptional Activation; D014170:Transformation, Genetic; D016335:Zinc Fingers
chemical_list: D029681:Arabidopsis Proteins; C518800:RAD54 protein, Arabidopsis; D011994:Recombinant Proteins; D003851:Deoxyribonucleases; D004265:DNA Helicases