Brief Communication

Plant Molecular Biology

, Volume 83, Issue 3, pp 279-285

TAL effector nucleases induce mutations at a pre-selected location in the genome of primary barley transformants

  • Toni WendtAffiliated withResearch Centre Flakkebjerg, Department of Molecular Biology and Genetics, Aarhus University Email author 
  • , Preben Bach HolmAffiliated withResearch Centre Flakkebjerg, Department of Molecular Biology and Genetics, Aarhus University
  • , Colby G. StarkerAffiliated withDepartment of Genetics, Cell Biology and Development and Center for Genome Engineering, University of Minnesota
  • , Michelle ChristianAffiliated withDepartment of Genetics, Cell Biology and Development and Center for Genome Engineering, University of Minnesota
  • , Daniel F. VoytasAffiliated withDepartment of Genetics, Cell Biology and Development and Center for Genome Engineering, University of Minnesota
  • , Henrik Brinch-PedersenAffiliated withResearch Centre Flakkebjerg, Department of Molecular Biology and Genetics, Aarhus University
  • , Inger Bæksted HolmeAffiliated withResearch Centre Flakkebjerg, Department of Molecular Biology and Genetics, Aarhus University

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.

Keywords

TAL effector nucleases Targeted mutagenesis Hordeum vulgare Cereal transformation TALEN