What is a repositorium?

The repositorium is a searchable database that provides data on relevant articles from journals, company web pages and web pages of governmental agencies about studies/applications of genome-editing in model plants and agricultural crops in the period January 1996 to May 2018. Search options are article type, technique, plant, traits or free text. The repositorium is based on the systematic map of Dominik Modrzejewski et al., published in the journal environmental evidence. (Download article PDF).

High frequency targeted mutagenesis in Arabidopsis thaliana using zinc finger nucleases

Typ / Jahr

Journal Article / 2010


Zhang, Feng; Maeder, Morgan L.; Unger-Wallace, Erica; Hoshaw, Justin P.; Reyon, Deepak; Christian, Michelle; Li, Xiaohong; Pierick, Christopher J.; Dobbs, Drena; Peterson, Thomas; Joung, J. Keith; Voytas, Daniel F.


We report here an efficient method for targeted mutagenesis of Arabidopsis genes through regulated expression of zinc finger nucleases (ZFNs)-enzymes engineered to create DNA double-strand breaks at specific target loci. ZFNs recognizing the Arabidopsis ADH1 and TT4 genes were made by Oligomerized Pool ENgineering (OPEN)-a publicly available, selection-based platform that yields high quality zinc finger arrays. The ADH1 and TT4 ZFNs were placed under control of an estrogen-inducible promoter and introduced into Arabidopsis plants by floral-dip transformation. Primary transgenic Arabidopsis seedlings induced to express the ADH1 or TT4 ZFNs exhibited somatic mutation frequencies of 7% or 16%, respectively. The induced mutations were typically insertions or deletions (1-142 bp) that were localized at the ZFN cleavage site and likely derived from imprecise repair of chromosome breaks by nonhomologous end-joining. Mutations were transmitted to the next generation for 69% of primary transgenics expressing the ADH1 ZFNs and 33% of transgenics expressing the TT4 ZFNs. Furthermore, approximately 20% of the mutant-producing plants were homozygous for mutations at ADH1 or TT4, indicating that both alleles were disrupted. ADH1 and TT4 were chosen as targets for this study because of their selectable or screenable phenotypes (adh1, allyl alcohol resistance; tt4, lack of anthocyanins in the seed coat). However, the high frequency of observed ZFN-induced mutagenesis suggests that targeted mutations can readily be recovered by simply screening progeny of primary transgenic plants by PCR and DNA sequencing. Taken together, our results suggest that it should now be possible to obtain mutations in any Arabidopsis target gene regardless of its mutant phenotype.

Alcohol Dehydrogenase/genetics; Arabidopsis Proteins/genetics; Arabidopsis/genetics/metabolism; Base Sequence; Deoxyribonucleases/genetics/metabolism; DNA repair; DNA, Plant/genetics/metabolism; gene targeting; Genes, Plant; Molecular Sequence Data; Mutagenesis, Site-Directed; Plants, Genetically Modified; Protein Engineering; Protoplasts/metabolism; Saccharomyces cerevisiae/genetics/metabolism; Zinc Fingers/genetics
Proceedings of the National Academy of Sciences of the United States of America
Periodical Number
Page range


ID Corresponding Author
Plant Species GE Technique
Sequence Identifier
Type of Alteration
Progress in Research
Key Topic
601 Voytas, Daniel F.
Arabidopsis thaliana Zinc-finger nucleases
Allyl alcohol resistance
Basic research
Basic research
602 Voytas, Daniel F.
Arabidopsis thaliana Zinc-finger nucleases
No information
Basic research
Basic research