The genetic intractability of Symbiodinium microadriaticum to standard algal transformation methods.
Journal
PloS one
ISSN: 1932-6203
Titre abrégé: PLoS One
Pays: United States
ID NLM: 101285081
Informations de publication
Date de publication:
2019
2019
Historique:
received:
06
06
2018
accepted:
24
01
2019
entrez:
20
2
2019
pubmed:
20
2
2019
medline:
5
11
2019
Statut:
epublish
Résumé
Modern transformation and genome editing techniques have shown great success across a broad variety of organisms. However, no study of successfully applied genome editing has been reported in a dinoflagellate despite the first genetic transformation of Symbiodinium being published about 20 years ago. Using an array of different available transformation techniques, we attempted to transform Symbiodinium microadriaticum (CCMP2467), a dinoflagellate symbiont of reef-building corals, with the view to performing subsequent CRISPR-Cas9 mediated genome editing. Plasmid vectors designed for nuclear transformation containing the chloramphenicol resistance gene under the control of the CaMV p35S promoter as well as several putative endogenous promoters were used to test a variety of transformation techniques including biolistics, electroporation and agitation with silicon carbide whiskers. Chloroplast-targeted transformation was attempted using an engineered Symbiodinium chloroplast minicircle encoding a modified PsbA protein expected to confer atrazine resistance. We report that we have been unable to confer chloramphenicol or atrazine resistance on Symbiodinium microadriaticum strain CCMP2467.
Identifiants
pubmed: 30779749
doi: 10.1371/journal.pone.0211936
pii: PONE-D-18-16942
pmc: PMC6380556
doi:
Substances chimiques
Chloroplast Proteins
0
Chloramphenicol
66974FR9Q1
Atrazine
QJA9M5H4IM
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e0211936Déclaration de conflit d'intérêts
The authors have declared that no competing interests exist.
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