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).

Increasing cloning possibilities using artificial zinc finger nucleases


Typ / Jahr

Journal Article / 2008

Autoren

Zeevi, Vardit; Tovkach, Andriy; Tzfira, Tzvi

Abstract

The ability to accurately digest and ligate DNA molecules of different origins is fundamental to modern recombinant DNA research. Only a handful of enzymes are capable of recognizing and cleaving novel and long DNA sequences, however. The slow evolution and engineering of new restriction enzymes calls for alternative strategies to design novel and unique restriction enzymes capable of binding and digesting specific long DNA sequences. Here we report on the use of zinc finger nucleases (ZFNs)-hybrid synthetic restriction enzymes that can be specifically designed to bind and cleave long DNA sequences-for the purpose of DNA recombination. We show that novel ZFNs can be designed for the digestion of specific sequences and can be expressed and used for cloning purposes. We also demonstrate the power of ZFNs in DNA cloning by custom-cloning a target DNA sequence and assembling dual-expression cassettes on a single target plasmid, a task that rarely can be achieved using type-II restriction enzymes. We demonstrate the flexibility of ZFN design and the ability to shuffle monomers of different ZFNs for the digestion of compatible recognition sites through ligation of compatible ends and their cleavage by heterodimer ZFNs. Of no less importance, we show that ZFNs can be designed to recognize and cleave existing DNA sequences for the custom-cloning of native target DNA molecules.

Keywords
Amino Acid Sequence; Arabidopsis/genetics; Base Sequence; Cloning, Molecular; DNA/metabolism; Endonucleases/chemistry/metabolism; Molecular Sequence Data; Plasmids/genetics; Zinc Fingers
Periodical
Proceedings of the National Academy of Sciences of the United States of America
Periodical Number
35
Page range
12785–12790
Volume
105
DOI
10.1073/pnas.0803618105

Techniques

ID Corresponding Author
Country
Plant Species GE Technique
Sequence Identifier
Trait
Type of Alteration
Progress in Research
Key Topic
1192 Tzfira, Tzvi
Israel
Arabidopsis thaliana Zinc-finger nucleases
H2a/H2b
No information
SDN1
Basic research
Basic research