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 Wendt
  • Preben Bach Holm
  • Colby G. Starker
  • Michelle Christian
  • Daniel F. Voytas
  • Henrik Brinch-Pedersen
  • Inger Bæksted Holme
Brief Communication

DOI: 10.1007/s11103-013-0078-4

Cite this article as:
Wendt, T., Holm, P.B., Starker, C.G. et al. Plant Mol Biol (2013) 83: 279. doi:10.1007/s11103-013-0078-4

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 nucleasesTargeted mutagenesisHordeum vulgareCereal transformationTALEN

Supplementary material

11103_2013_78_MOESM1_ESM.pdf (89 kb)
Supplementary material 1 (PDF 88 kb)

Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Toni Wendt
    • 1
  • Preben Bach Holm
    • 1
  • Colby G. Starker
    • 2
  • Michelle Christian
    • 2
  • Daniel F. Voytas
    • 2
  • Henrik Brinch-Pedersen
    • 1
  • Inger Bæksted Holme
    • 1
  1. 1.Research Centre Flakkebjerg, Department of Molecular Biology and GeneticsAarhus UniversitySlagelseDenmark
  2. 2.Department of Genetics, Cell Biology and Development and Center for Genome EngineeringUniversity of MinnesotaMinneapolisUSA