Repositorium

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).

Rapid and highly efficient construction of TALE-based transcriptional regulators and nucleases for genome modification


Typ / Jahr

Journal Article / 2012

Autoren

Li, Lixin; Piatek, Marek J.; Atef, Ahmed; Piatek, Agnieszka; Wibowo, Anjar; Fang, Xiaoyun; Sabir, J. S. M.; Zhu, Jian-Kang; Mahfouz, Magdy M.

Abstract

Transcription activator-like effectors (TALEs) can be used as DNA-targeting modules by engineering their repeat domains to dictate user-selected sequence specificity. TALEs have been shown to function as site-specific transcriptional activators in a variety of cell types and organisms. TALE nucleases (TALENs), generated by fusing the FokI cleavage domain to TALE, have been used to create genomic double-strand breaks. The identity of the TALE repeat variable di-residues, their number, and their order dictate the DNA sequence specificity. Because TALE repeats are nearly identical, their assembly by cloning or even by synthesis is challenging and time consuming. Here, we report the development and use of a rapid and straightforward approach for the construction of designer TALE (dTALE) activators and nucleases with user-selected DNA target specificity. Using our plasmid set of 100 repeat modules, researchers can assemble repeat domains for any 14-nucleotide target sequence in one sequential restriction-ligation cloning step and in only 24 h. We generated several custom dTALEs and dTALENs with new target sequence specificities and validated their function by transient expression in tobacco leaves and in vitro DNA cleavage assays, respectively. Moreover, we developed a web tool, called idTALE, to facilitate the design of dTALENs and the identification of their genomic targets and potential off-targets in the genomes of several model species. Our dTALE repeat assembly approach along with the web tool idTALE will expedite genome-engineering applications in a variety of cell types and organisms including plants.

Keywords
Deoxyribonucleases, Type II Site-Specific; DNA Breaks, Double-Stranded; DNA cleavage; DNA-Binding Proteins/genetics/metabolism; Endonucleases/genetics/metabolism; Homeodomain Proteins/genetics; Plant Leaves/genetics; Protein Engineering/methods; Recombinant Proteins/genetics/metabolism; Repetitive Sequences, Nucleic Acid; Reproducibility of Results; Tobacco/genetics; Trans-Activators/genetics
Periodical
Plant molecular biology
Periodical Number
4-5
Page range
407–416
Volume
78
DOI
10.1007/s11103-012-9875-4

Techniques

ID Corresponding Author
Country
Plant Species GE Technique
Sequence Identifier
Trait
Type of Alteration
Progress in Research
Key Topic
227 Mahfouz Magdy M.; Zhu, Jian-Kang
Saudi Arabia, USA
Arabidopsis thaliana TALENs
PDS3
Albino and dwarf phenotype
SDN1
Basic research
Basic research