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).

Genotyping genome-edited mutations in plants using CRISPR ribonucleoprotein complexes


Typ / Jahr

Journal Article / 2018

Autoren

Liang, Zhen; Chen, Kunling; Yan, Yan; Zhang, Yi; Gao, Caixia

Abstract

Despite the great achievements in genome editing, accurately detecting mutations induced by sequence-specific nucleases is still a challenge in plants, especially in polyploidy plants. An efficient detection method is particularly vital when the mutation frequency is low or when a large population needs to be screened. Here, we applied purified CRISPR ribonucleoprotein complexes to cleave PCR products for genome-edited mutation detection in hexaploid wheat and diploid rice. We show that this mutation detection method is more sensitive than Sanger sequencing and more applicable than PCR/RE method without the requirement for restriction enzyme site. We also demonstrate that this detection method is especially useful for genome editing in wheat, because target sites are often surrounded by single nucleotide polymorphisms. Using this screening method, we were also able to detect foreign DNA-free tagw2 mutations induced by purified TALEN protein. Finally, we show that partial base editing mutations can also be detected using high-fidelity SpCas9 variants or FnCpf1. The PCR/RNP method is low-cost and widely applicable for rapid detection of genome-edited mutation in plants.

Keywords
FnCpf1; hexaploid wheat; high-fidelity SpCas9 variants.; PCR/RNP; SpCas9; TALEN protein
Periodical
Plant Biotechnol J (Plant Biotechnology Journal)
Periodical Number
Page range
Volume
42
DOI
10.1111/pbi.12938

Techniques

ID Corresponding Author
Country
Plant Species GE Technique
Sequence Identifier
Trait
Type of Alteration
Progress in Research
Key Topic
748 Gao, Caixia
China
Oryza sativa CRISPR/Cas9
Cer9
No information
SDN1
Basic research
Basic research
749 Gao, Caixia
China
Oryza sativa CRISPR/Cas9
PDS
Albino phenotype
SDN1
Basic research
Basic research
750 Gao, Caixia
China
Oryza sativa Base-editing
PDS1
Albino phenotype
BE
Basic research
Basic research
751 Gao, Caixia
China
Oryza sativa Base-editing
PDS4
Albino phenotype
BE
Basic research
Basic research
752 Gao, Caixia
China
Triticum aestivum CRISPR/Cas9
GW2
Increase in seed size and thousand grain weight
SDN1
Basic research
Basic research
753 Gao, Caixia
China
Triticum aestivum CRISPR/Cpf1
GW2
Increase in seed size and thousand grain weight
SDN1
Basic research
Basic research
754 Gao, Caixia
China
Triticum aestivum TALENs
GW2
Increase in seed size and thousand grain weight
SDN1
Basic research
Basic research
755 Gao, Caixia
China
Triticum aestivum CRISPR/Cas9
Cer9
No information
SDN1
Basic research
Basic research
756 Gao, Caixia
China
Triticum aestivum CRISPR/Cas9
GASR7
No information
SDN1
Basic research
Basic research
757 Gao, Caixia
China
Triticum aestivum CRISPR/Cpf1
GASR7
No information
SDN1
Basic research
Basic research