A Universal System of CRISPR/Cas9-Mediated Gene Targeting Using All-in-One Vector in Plants.
CRISPR/Cas9
gene targeting
genome editing
homologous recombination
rice
Journal
Frontiers in genome editing
ISSN: 2673-3439
Titre abrégé: Front Genome Ed
Pays: Switzerland
ID NLM: 101775540
Informations de publication
Date de publication:
2020
2020
Historique:
received:
09
09
2020
accepted:
19
10
2020
entrez:
29
10
2021
pubmed:
30
10
2021
medline:
30
10
2021
Statut:
epublish
Résumé
Homologous recombination-mediated genome editing, also called gene targeting (GT), is an essential technique that allows precise modification of a target sequence, including introduction of point mutations, knock-in of a reporter gene, and/or swapping of a functional domain. However, due to its low frequency, it has been difficult to establish GT approaches that can be applied widely to a large number of plant species. We have developed a simple and universal clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-mediated DNA double-strand break (DSB)-induced GT system using an all-in-one vector comprising a CRISPR/Cas9 expression construct, selectable marker, and GT donor template. This system enabled introduction of targeted point mutations with non-selectable traits into several target genes in both rice and tobacco. Since it was possible to evaluate the GT frequency on endogenous target genes precisely using this system, we investigated the effect of treatment with Rad51-stimulatory compound 1 (RS-1) on the frequency of DSB-induced GT. GT frequency was slightly, but consistently, improved by RS-1 treatment in both target plants.
Identifiants
pubmed: 34713227
doi: 10.3389/fgeed.2020.604289
pmc: PMC8525384
doi:
Types de publication
Journal Article
Langues
eng
Pagination
604289Informations de copyright
Copyright © 2020 Nishizawa-Yokoi, Mikami and Toki.
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