Creating Targeted Gene Knockouts in Brassica oleracea Using CRISPR/Cas9.
Brassica oleracea
CRISPR/Cas9
Gene edit
Genome editing
Knockout
Mutant
Transgene free
Vector
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2019
2019
Historique:
entrez:
6
1
2019
pubmed:
6
1
2019
medline:
18
6
2019
Statut:
ppublish
Résumé
While public and political views on genetic modification (inserting "foreign" genes to elicit new traits) have resulted in limited exploitation of the technology in some parts of the world, the new era of genome editing (to edit existing genes to gain new traits/genetic variation) has the potential to change the biotech landscape. Genome editing offers a faster and simpler approach to gene knockout in both single and multiple genetic locations, within a single or small number of generations, in a way that has not been possible through alternative breeding methods. Here we describe an Agrobacterium-mediated delivery approach to deliver Cas9 and dual sgRNAs into 4-day-old cotyledonary petioles of Brassica oleracea. Mutations are detected in approximately 10% of primary transgenic plants and go on in subsequent T1 and T2 generations to segregate away from the T-DNA. This enables the recovery of non-transgenic, genome-edited plants carrying a variety of mutations at the target locus.
Identifiants
pubmed: 30610635
doi: 10.1007/978-1-4939-8991-1_12
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
155-170Subventions
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/N019466/1
Pays : United Kingdom
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/P013511/1
Pays : United Kingdom
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