Targeted Mutagenesis of Arabidopsis thaliana Using Engineered TAL Effector Nucleases

Michelle Christian, Yiping Qi, Yong Zhang, Daniel F. Voytas

Research output: Contribution to journalArticlepeer-review

85 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1697-1705
Number of pages9
JournalG3: Genes, Genomes, Genetics
Volume3
Issue number9
DOIs
StatePublished - 2013

Keywords

  • Arabidopsis
  • End-joining
  • Mutagenesis
  • Nonhomologous
  • Nuclease
  • TAL effector
  • Targeted
  • Thaliana

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