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).

Zinc finger nuclease-mediated transgene deletion


Typ / Jahr

Journal Article / 2010

Autoren

Petolino, Joseph F.; Worden, Andrew; Curlee, Krisi; Connell, James; Strange Moynahan, Tonya L.; Larsen, Cory; Russell, Sean

Abstract

A transgene, flanked by zinc finger nuclease (ZFN) cleavage sites, was deleted from a stably transformed plant by crossing it with a second plant expressing a corresponding ZFN gene. A target construct, containing a GUS reporter gene flanked by ZFN cleavage sites, a GFP reporter gene and a PAT selectable marker gene, was transformed into tobacco. Basta-resistant plants were regenerated and screened for GUS and GFP expression. A second construct, containing a ZFN gene driven by the constitutive CsVMV promoter and an HPT selectable marker gene, was also transformed into tobacco. Selected T(0) plants were grown to maturity and allowed to self-pollinate. Homozygous target plants, which expressed GUS and GFP, were crossed with homozygous ZFN plants, which expressed the ZFN gene. Numerous GUS-negative plants were observed among the hybrids with one particular cross displaying approximately 35% GUS-negative plants. Evidence for complete deletion of a 4.3 kb sequence comprising the GUS gene was obtained and sequence confirmed. Co-segregation in F(2) progenies of 'truncated' and 'intact' target sequences with expected reporter gene phenotypes were observed. Since ZFNs can be designed to bind and cleave a wide range of DNA sequences, these results constitute a general strategy for creating targeted gene deletions.

Keywords
Base Sequence; Blotting, Southern; Crosses, Genetic; Endonucleases/genetics/metabolism; Glucuronidase/genetics/metabolism; Green Fluorescent Proteins/genetics/metabolism; Molecular Sequence Data; Plants, Genetically Modified/genetics/metabolism; Sequence Deletion/genetics; Sequence Homology, Nucleic Acid; Tobacco/genetics/metabolism; Transgenes/genetics; Zinc Fingers
Periodical
Plant molecular biology
Periodical Number
6
Page range
617–628
Volume
73
DOI
10.1007/s11103-010-9641-4

Techniques

ID Corresponding Author
Country
Plant Species GE Technique
Sequence Identifier
Trait
Type of Alteration
Progress in Research
Key Topic
363 Petolino, Joseph F.
USA
Tobacco Zinc-finger nucleases
integrated ZFN target sequence
No information
SDN1
Basic research
Basic research