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).

Development of germ-line-specific CRISPR-Cas9 systems to improve the production of heritable gene modifications in Arabidopsis


Typ / Jahr

Journal Article / 2016

Autoren

Mao, Yanfei; Zhang, Zhengjing; Feng, Zhengyan; Wei, Pengliang; Zhang, Hui; Botella, José Ramón; Zhu, Jian-Kang

Abstract

The Streptococcus-derived CRISPR/Cas9 system is being widely used to perform targeted gene modifications in plants. This customized endonuclease system has two components, the single-guide RNA (sgRNA) for target DNA recognition and the CRISPR-associated protein 9 (Cas9) for DNA cleavage. Ubiquitously expressed CRISPR/Cas9 systems (UC) generate targeted gene modifications with high efficiency but only those produced in reproductive cells are transmitted to the next generation. We report the design and characterization of a germ-line-specific Cas9 system (GSC) for Arabidopsis gene modification in male gametocytes, constructed using a SPOROCYTELESS (SPL) genomic expression cassette. Four loci in two endogenous genes were targeted by both systems for comparative analysis. Mutations generated by the GSC system were rare in T1 plants but were abundant (30%) in the T2 generation. The vast majority (70%) of the T2 mutant population generated using the UC system were chimeras while the newly developed GSC system produced only 29% chimeras, with 70% of the T2 mutants being heterozygous. Analysis of two loci in the T2 population showed that the abundance of heritable gene mutations was 37% higher in the GSC system compared to the UC system and the level of polymorphism of the mutations was also dramatically increased with the GSC system. Two additional systems based on germ-line-specific promoters (pDD45-GT and pLAT52-GT) were also tested, and one of them was capable of generating heritable homozygous T1 mutant plants. Our results suggest that future application of the described GSC system will facilitate the screening for targeted gene modifications, especially lethal mutations in the T2 population.

Keywords
Arabidopsis/genetics; Base Sequence; chimera; Clustered Regularly Interspaced Short Palindromic Repeats/genetics; Crosses, Genetic; Genes, Plant; Genetic Engineering/methods; Genetic Vectors/metabolism; Genotyping Techniques; Germ Cells, Plant/metabolism; Germ Cells/metabolism; Hybridization, Genetic; Inheritance Patterns/genetics; Mutagenesis, Site-Directed; Mutation/genetics; Organ Specificity/genetics; Promoter Regions, Genetic/genetics
Periodical
Plant biotechnology journal
Periodical Number
2
Page range
519–532
Volume
14
DOI
10.1111/pbi.12468

Techniques

ID Corresponding Author
Country
Plant Species GE Technique
Sequence Identifier
Trait
Type of Alteration
Progress in Research
Key Topic
303 Zhu, Jian-Kang
China
Arabidopsis thaliana CRISPR/Cas9
AP1
No information
SDN1
Basic research
Basic research
304 Zhu, Jian-Kang
China
Arabidopsis thaliana CRISPR/Cas9
TT4
No information
SDN1
Basic research
Basic research