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).

Multigene knockout utilizing off-target mutations of the CRISPR/Cas9 system in rice


Typ / Jahr

Journal Article / 2015

Autoren

Endo, Masaki; Mikami, Masafumi; Toki, Seiichi

Abstract

The clustered regularly interspaced short palindromic repeat (CRISPR)-associated endonuclease 9 (CRISPR/Cas9) system has been demonstrated to be a robust genome engineering tool in a variety of organisms including plants. However, it has been shown that the CRISPR/Cas9 system cleaves genomic DNA sequences containing mismatches to the guide RNA strand. We expected that this low specificity could be exploited to induce multihomeologous and multiparalogous gene knockouts. In the case of polyploid plants, simultaneous modification of multiple homeologous genes, i.e. genes with similar but not identical DNA sequences, is often needed to obtain a desired phenotype. Even in diploid plants, disruption of multiparalogous genes, which have functional redundancy, is often needed. To validate the applicability of the CRISPR/Cas9 system to target mutagenesis of paralogous genes in rice, we designed a single-guide RNA (sgRNA) that recognized 20 bp sequences of cyclin-dependent kinase B2 (CDKB2) as an on-target locus. These 20 bp possess similarity to other rice CDK genes (CDKA1, CDKA2 and CDKB1) with different numbers of mismatches. We analyzed mutations in these four CDK genes in plants regenerated from Cas9/sgRNA-transformed calli and revealed that single, double and triple mutants of CDKA2, CDKB1 and CDKB2 can be created by a single sgRNA.

Keywords
Base Sequence; Clustered Regularly Interspaced Short Palindromic Repeats/genetics; CRISPR-Cas Systems; Cyclin-Dependent Kinases/genetics; Gene Knockout Techniques/methods; genetic engineering; mutagenesis; Mutation; Oryza/genetics; Plant Proteins/genetics; RNA, Guide/genetics; RNA, Plant/genetics
Periodical
Plant & cell physiology
Periodical Number
1
Page range
41–47
Volume
56
DOI
10.1093/pcp/pcu154

Techniques

ID Corresponding Author
Country
Plant Species GE Technique
Sequence Identifier
Trait
Type of Alteration
Progress in Research
Key Topic
98 Toki, Seiichi
Japan
Oryza sativa CRISPR/Cas9
CDKB2
No information
SDN1
Basic research
Basic research