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).

Transcription activator-like effector nucleases enable efficient plant genome engineering

Typ / Jahr

Journal Article / 2013


Zhang, Yong; Zhang, Feng; Li, Xiaohong; Baller, Joshua A.; Qi, Yiping; Starker, Colby G.; Bogdanove, Adam J.; Voytas, Daniel F.


The ability to precisely engineer plant genomes offers much potential for advancing basic and applied plant biology. Here, we describe methods for the targeted modification of plant genomes using transcription activator-like effector nucleases (TALENs). Methods were optimized using tobacco (Nicotiana tabacum) protoplasts and TALENs targeting the acetolactate synthase (ALS) gene. Optimal TALEN scaffolds were identified using a protoplast-based single-strand annealing assay in which TALEN cleavage creates a functional yellow fluorescent protein gene, enabling quantification of TALEN activity by flow cytometry. Single-strand annealing activity data for TALENs with different scaffolds correlated highly with their activity at endogenous targets, as measured by high-throughput DNA sequencing of polymerase chain reaction products encompassing the TALEN recognition sites. TALENs introduced targeted mutations in ALS in 30% of transformed cells, and the frequencies of targeted gene insertion approximated 14%. These efficiencies made it possible to recover genome modifications without selection or enrichment regimes: 32% of tobacco calli generated from protoplasts transformed with TALEN-encoding constructs had TALEN-induced mutations in ALS, and of 16 calli characterized in detail, all had mutations in one allele each of the duplicate ALS genes (SurA and SurB). In calli derived from cells treated with a TALEN and a 322-bp donor molecule differing by 6 bp from the ALS coding sequence, 4% showed evidence of targeted gene replacement. The optimized reagents implemented in plant protoplasts should be useful for targeted modification of cells from diverse plant species and using a variety of means for reagent delivery.

Acetolactate Synthase/genetics/metabolism; Alleles; Bacterial Proteins/metabolism; DNA, Plant/genetics; Endonucleases/metabolism; Gene Knockout Techniques; Genetic Engineering/methods; Genetic Vectors/genetics/metabolism; Genome, Plant; High-Throughput Nucleotide Sequencing; Luminescent Proteins/metabolism; Mutagenesis, Insertional/methods; Plant Cells/metabolism; Plasmids/genetics/metabolism; polymerase chain reaction; Protoplasts/cytology/metabolism; Tobacco/enzymology/genetics; Trans-Activators/metabolism; Transcriptional Activation; Transformation, Genetic
Plant physiology
Periodical Number
Page range


ID Corresponding Author
Plant Species GE Technique
Sequence Identifier
Type of Alteration
Progress in Research
Key Topic
632 Voytas, Daniel F.
Nicotiana tabacum TALENs
herbicide tolerance
Basic research
Basic research
633 Voytas, Daniel F.
Nicotiana tabacum TALENs
herbicide tolerance
Basic research
Basic research
634 Voytas, Daniel F.
Nicotiana tabacum TALENs
No information
YFP (Yellow fluorescent protein)
Basic research
Basic research