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).

High-frequency, precise modification of the tomato genome


Typ / Jahr

Journal Article / 2015

Autoren

Čermák, Tomáš; Baltes, Nicholas J.; Čegan, Radim; Zhang, Yong; Voytas, Daniel F.

Abstract

Background: The use of homologous recombination to precisely modify plant genomes has been challenging, due to the lack of efficient methods for delivering DNA repair templates to plant cells. Even with the advent of sequence-specific nucleases, which stimulate homologous recombination at predefined genomic sites by creating targeted DNA double-strand breaks, there are only a handful of studies that report precise editing of endogenous genes in crop plants. More efficient methods are needed to modify plant genomes through homologous recombination, ideally without randomly integrating foreign DNA. Results: Here, we use geminivirus replicons to create heritable modifications to the tomato genome at frequencies tenfold higher than traditional methods of DNA delivery (i.e., Agrobacterium). A strong promoter was inserted upstream of a gene controlling anthocyanin biosynthesis, resulting in overexpression and ectopic accumulation of pigments in tomato tissues. More than two-thirds of the insertions were precise, and had no unanticipated sequence modifications. Both TALENs and CRISPR/Cas9 achieved gene targeting at similar efficiencies. Further, the targeted modification was transmitted to progeny in a Mendelian fashion. Even though donor molecules were replicated in the vectors, no evidence was found of persistent extra-chromosomal replicons or off-target integration of T-DNA or replicon sequences. Conclusions: High-frequency, precise modification of the tomato genome was achieved using geminivirus replicons, suggesting that these vectors can overcome the efficiency barrier that has made gene targeting in plants challenging. This work provides a foundation for efficient genome editing of crop genomes without the random integration of foreign DNA.

Keywords
Anthocyanins/biosynthesis/genetics; CRISPR-Cas Systems/genetics; DNA Breaks, Double-Stranded; DNA Repair/genetics; DNA, Bacterial/genetics; Geminiviridae/genetics; gene targeting; genetic engineering; Genome, Plant; Homologous Recombination/genetics; Lycopersicon esculentum/genetics
Periodical
Genome biology
Periodical Number
Page range
232
Volume
16
DOI
10.1186/s13059-015-0796-9

Techniques

ID Corresponding Author
Country
Plant Species GE Technique
Sequence Identifier
Trait
Type of Alteration
Progress in Research
Key Topic
46 Voytas, Daniel F.
USA
Solanum lycopersicum CRISPR/Cas9
ANT1
anthocyan accumulation/ purple plant tissue
SDN1
Basic research
Basic research
47 Voytas, Daniel F.
USA
Solanum lycopersicum TALENs
ANT1
anthocyan accumulation/ purple plant tissue
SDN1
Basic research
Basic research