title: Targeted mutagenesis of Arabidopsis thaliana using engineered TAL effector nucleases.
abstract: Custom TAL effector nucleases (TALENs) are increasingly used as reagents to manipulate genomes in vivo. Here, we used TALENs to modify the genome of the model plant, Arabidopsis thaliana. We engineered seven TALENs targeting five Arabidopsis genes, namely ADH1, TT4, MAPKKK1, DSK2B, and NATA2. In pooled seedlings expressing the TALENs, we observed somatic mutagenesis frequencies ranging from 2-15% at the intended targets for all seven TALENs. Somatic mutagenesis frequencies as high as 41-73% were observed in individual transgenic plant lines expressing the TALENs. Additionally, a TALEN pair targeting a tandemly duplicated gene induced a 4.4-kb deletion in somatic cells. For the most active TALEN pairs, namely those targeting ADH1 and NATA2, we found that TALEN-induced mutations were transmitted to the next generation at frequencies of 1.5-12%. Our work demonstrates that TALENs are useful reagents for achieving targeted mutagenesis in this important plant model.
authors: Department of Genetics, Cell Biology, and Development and Center for Genome Engineering, University of Minnesota, Minneapolis, Minnesota 55455.
authors: Michelle
authors: M
authors: Christian
authors: Yiping
authors: Y
authors: Qi
authors: Yong
authors: Y
authors: Zhang
authors: Daniel F
authors: DF
authors: Voytas
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000235:genetics*; D001483:Base Sequence; D004262:DNA Restriction Enzymes / Q000235:genetics*; D018390:Gene Targeting / Q000379:methods*; D017343:Genes, Plant; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D030821:Plants, Genetically Modified
chemical_list: D004262:DNA Restriction Enzymes
keywords: Arabidopsis thaliana; TAL effector nuclease; nonhomologous end-joining; targeted mutagenesis
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