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).

Induction of targeted, heritable mutations in barley and Brassica oleracea using RNA-guided Cas9 nuclease


Typ / Jahr

Journal Article / 2015

Autoren

Lawrenson, Tom; Shorinola, Oluwaseyi; Stacey, Nicola; Li, Chengdao; Østergaard, Lars; Patron, Nicola; Uauy, Cristobal; Harwood, Wendy

Abstract

Background: The RNA-guided Cas9 system represents a flexible approach for genome editing in plants. This method can create specific mutations that knock-out or alter target gene function. It provides a valuable tool for plant research and offers opportunities for crop improvement. Results: We investigate the use and target specificity requirements of RNA-guided Cas9 genome editing in barley (Hordeum vulgare) and Brassica oleracea by targeting multicopy genes. In barley, we target two copies of HvPM19 and observe Cas9-induced mutations in the first generation of 23 % and 10 % of the lines, respectively. In B. oleracea, targeting of BolC.GA4.a leads to Cas9-induced mutations in 10 % of first generation plants screened. In addition, a phenotypic screen identifies T0 plants with the expected dwarf phenotype associated with knock-out of the target gene. In both barley and B. oleracea stable Cas9-induced mutations are transmitted to T2 plants independently of the T-DNA construct. We observe off-target activity in both species, despite the presence of at least one mismatch between the single guide RNA and the non-target gene sequences. In barley, a transgene-free plant has concurrent mutations in the target and non-target copies of HvPM19. Conclusions: We demonstrate the use of RNA-guided Cas9 to generate mutations in target genes of both barley and B. oleracea and show stable transmission of these mutations thus establishing the potential for rapid characterisation of gene function in these species. In addition, the off-target effects reported offer both potential difficulties and specific opportunities to target members of multigene families in crops.

Keywords
Barley; Brassica; breeding; CRISPR/Cas9; Crops; GA4; genome editing; Mutations; Off-target; PM19
Periodical
Genome biology
Periodical Number
Page range
258
Volume
16
DOI
10.1186/s13059-015-0826-7

Techniques

ID Corresponding Author
Country
Plant Species GE Technique
Sequence Identifier
Trait
Type of Alteration
Progress in Research
Key Topic
213 Uauy, Cristobal; Harwood, Wendy
United Kingdom
Hordeum vulgare CRISPR/Cas9
PM19
Positive regulator of grain dormancy
SDN1
Basic research
Basic research
214 Uauy, Cristobal; Harwood, Wendy
United Kingdom
Brassica oleracea CRISPR/Cas9
GA4a
Involved in pod valve margin development
SDN1
Basic research
Basic research