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).

Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting

26

Typ / Jahr

Journal Article / 2011

Autoren

Cermak, T.; Doyle, E. L.; Christian, M.; Wang, L.; Zhang, Y.; Schmidt, C.; Baller, J. A.; Somia, N. V.; Bogdanove, A. J.; Voytas, D. F.

Abstract

TALENs are important new tools for genome engineering. Fusions of transcription activator-like (TAL) effectors of plant pathogenic Xanthomonas spp. to the FokI nuclease, TALENs bind and cleave DNA in pairs. Binding specificity is determined by customizable arrays of polymorphic amino acid repeats in the TAL effectors. We present a method and reagents for efficiently assembling TALEN constructs with custom repeat arrays. We also describe design guidelines based on naturally occurring TAL effectors and their binding sites. Using software that applies these guidelines, in nine genes from plants, animals and protists, we found candidate cleavage sites on average every 35 bp. Each of 15 sites selected from this set was cleaved in a yeast-based assay with TALEN pairs constructed with our reagents. We used two of the TALEN pairs to mutate HPRT1 in human cells and ADH1 in Arabidopsis thaliana protoplasts. Our reagents include a plasmid construct for making custom TAL effectors and one for TAL effector fusions to additional proteins of interest. Using the former, we constructed de novo a functional analog of AvrHah1 of Xanthomonas gardneri. The complete plasmid set is available through the non-profit repository AddGene and a web-based version of our software is freely accessible online.

Keywords

Amino Acid Sequence; Arabidopsis/genetics; Base Sequence; Deoxyribonucleases, Type II Site-Specific/chemistry/genetics/metabolism; DNA cleavage; DNA-Binding Proteins/chemistry/genetics/metabolism; gene targeting; Humans; Molecular Sequence Data; mutagenesis; Protein Engineering/methods; Protoplasts/metabolism; Recombinant Fusion Proteins/chemistry/metabolism; Repetitive Sequences, Amino Acid; Software; Trans-Activators/chemistry/genetics/metabolism; Xanthomonas/genetics

Periodical

Nucleic acids research

Periodical Number

Page range

Volume

DOI

10.1093/nar/gkr218

View Publication

Techniques

ID	Corresponding Author Country	Plant Species	GE Technique Sequence Identifier	Trait Type of Alteration	Progress in Research Key Topic
45	Voytas, Daniel F. USA	Arabidopsis thaliana	TALENs ADH1	Allyl alcohol resistance SDN1	Basic research Basic research