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).

Selection of highly efficient sgRNAs for CRISPR/Cas9-based plant genome editing


Typ / Jahr

Journal Article / 2016

Autoren

Liang, Gang; Zhang, Huimin; Lou, Dengji; Yu, Diqiu

Abstract

The CRISPR/Cas9-sgRNA system has been developed to mediate genome editing and become a powerful tool for biological research. Employing the CRISPR/Cas9-sgRNA system for genome editing and manipulation has accelerated research and expanded researchers’ ability to generate genetic models. However, the method evaluating the efficiency of sgRNAs is lacking in plants. Based on the nucleotide compositions and secondary structures of sgRNAs which have been experimentally validated in plants, we instituted criteria to design efficient sgRNAs. To facilitate the assembly of multiple sgRNA cassettes, we also developed a new strategy to rapidly construct CRISPR/Cas9-sgRNA system for multiplex editing in plants. In theory, up to ten single guide RNA (sgRNA) cassettes can be simultaneously assembled into the final binary vectors. As a proof of concept, 21 sgRNAs complying with the criteria were designed and the corresponding Cas9/sgRNAs expression vectors were constructed. Sequencing analysis of transgenic rice plants suggested that 82% of the desired target sites were edited with deletion, insertion, substitution, and inversion, displaying high editing efficiency. This work provides a convenient approach to select efficient sgRNAs for target editing.

Keywords
Base Sequence; CRISPR-Cas Systems; gene editing; Gene Order; Genetic Vectors/genetics; Genome, Plant; Genotype; Position-Specific Scoring Matrices; RNA, Guide/chemistry/genetics; RNA, Plant/chemistry/genetics
Periodical
Scientific reports
Periodical Number
Page range
21451
Volume
6
DOI
10.1038/srep21451

Techniques

ID Corresponding Author
Country
Plant Species GE Technique
Sequence Identifier
Trait
Type of Alteration
Progress in Research
Key Topic
242 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os07g35870
No information
SDN1
Basic research
Basic research
243 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os08g04390
No information
SDN1
Basic research
Basic research
244 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os03g27280
No information
SDN1
Basic research
Basic research
245 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os10g41490
No information
SDN1
Basic research
Basic research
246 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os05g38140
No information
SDN1
Basic research
Basic research
247 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os02g34600
No information
SDN1
Basic research
Basic research
248 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os12g39630
No information
SDN1
Basic research
Basic research
249 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os03g55600
No information
SDN1
Basic research
Basic research
250 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os04g35240
No information
SDN1
Basic research
Basic research
251 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os04g31290
No information
SDN1
Basic research
Basic research
252 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os02g02480
No information
SDN1
Basic research
Basic research
253 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os07g35870
No information
SDN1
Basic research
Basic research
254 Yu, Diqiu
China
Oryza sativa CRISPR/Cas9
Os02g47220
No information
SDN1
Basic research
Basic research