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).

Targeted mutagenesis of duplicated genes in soybean with zinc-finger nucleases


Typ / Jahr

Journal Article / 2011

Autoren

Curtin, Shaun J.; Zhang, Feng; Sander, Jeffry D.; Haun, William J.; Starker, Colby; Baltes, Nicholas J.; Reyon, Deepak; Dahlborg, Elizabeth J.; Goodwin, Mathew J.; Coffman, Andrew P.; Dobbs, Drena; Joung, J. Keith; Voytas, Daniel F.; Stupar, Robert M.

Abstract

We performed targeted mutagenesis of a transgene and nine endogenous soybean (Glycine max) genes using zinc-finger nucleases (ZFNs). A suite of ZFNs were engineered by the recently described context-dependent assembly platform--a rapid, open-source method for generating zinc-finger arrays. Specific ZFNs targeting dicer-like (DCL) genes and other genes involved in RNA silencing were cloned into a vector under an estrogen-inducible promoter. A hairy-root transformation system was employed to investigate the efficiency of ZFN mutagenesis at each target locus. Transgenic roots exhibited somatic mutations localized at the ZFN target sites for seven out of nine targeted genes. We next introduced a ZFN into soybean via whole-plant transformation and generated independent mutations in the paralogous genes DCL4a and DCL4b. The dcl4b mutation showed efficient heritable transmission of the ZFN-induced mutation in the subsequent generation. These findings indicate that ZFN-based mutagenesis provides an efficient method for making mutations in duplicate genes that are otherwise difficult to study due to redundancy. We also developed a publicly accessible Web-based tool to identify sites suitable for engineering context-dependent assembly ZFNs in the soybean genome.

Keywords
Base Sequence; Endonucleases/chemistry/metabolism; Genes, Duplicate/genetics; Genes, Plant/genetics; Genetic Techniques; Green Fluorescent Proteins/metabolism; Inheritance Patterns/genetics; Internet; Molecular Sequence Data; Mutagenesis/genetics; Mutation/genetics; Plant Roots/genetics; polymerase chain reaction; Soybeans/genetics; Transgenes/genetics; Zinc Fingers/genetics
Periodical
Plant physiology
Periodical Number
2
Page range
466-473
Volume
156
DOI
10.1104/pp.111.172981

Techniques

ID Corresponding Author
Country
Plant Species GE Technique
Sequence Identifier
Trait
Type of Alteration
Progress in Research
Key Topic
76 Stupar, Robert M.
USA
Glycine Max Zinc-finger nucleases
DCL
strong morphological phenotype, characterized by reduced seed size and aborted seedling development, in addition to defective miRNA precursor transcript processing efficiency and deregulated miRNA target gene expression
SDN1
Basic research
Basic research
77 Stupar, Robert M.
USA
Glycine Max Zinc-finger nucleases
HEN
No information
SDN1
Basic research
Basic research
78 Stupar, Robert M.
USA
Glycine Max Zinc-finger nucleases
RDR
No information
SDN1
Basic research
Basic research