title: TAL effector nucleases induce mutations at a pre-selected location in the genome of primary barley transformants.
abstract: Transcription activator-like effector nucleases (TALENs) enable targeted mutagenesis in a variety of organisms. The primary advantage of TALENs over other sequence-specific nucleases, namely zinc finger nucleases and meganucleases, lies in their ease of assembly, reliability of function, and their broad targeting range. Here we report the assembly of several TALENs for a specific genomic locus in barley. The cleavage activity of individual TALENs was first tested in vivo using a yeast-based, single-strand annealing assay. The most efficient TALEN was then selected for barley transformation. Analysis of the resulting transformants showed that TALEN-induced double strand breaks led to the introduction of short deletions at the target site. Additional analysis revealed that each barley transformant contained a range of different mutations, indicating that mutations occurred independently in different cells.
authors.affiliation: Research Centre Flakkebjerg, Department of Molecular Biology and Genetics, Aarhus University, Slagelse, Denmark, toni.wendt@agrsci.dk.
authors.forename: Toni
authors.initials: T
authors.lastname: Wendt
authors.forename: Preben Bach
authors.initials: PB
authors.lastname: Holm
authors.forename: Colby G
authors.initials: CG
authors.lastname: Starker
authors.forename: Michelle
authors.initials: M
authors.lastname: Christian
authors.forename: Daniel F
authors.initials: DF
authors.lastname: Voytas
authors.forename: Henrik
authors.initials: H
authors.lastname: Brinch-Pedersen
authors.forename: Inger Bæksted
authors.initials: IB
authors.lastname: Holme
mesh_terms: D001483:Base Sequence; D004720:Endonucleases / Q000378:metabolism*; D018745:Genome, Plant; D001467:Hordeum / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D012689:Sequence Homology, Nucleic Acid; D014157:Transcription Factors / Q000378:metabolism*; D014170:Transformation, Genetic
chemical_list: D014157:Transcription Factors; D004720:Endonucleases