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The genetic intractability of Symbiodinium microadriaticum to standard algal transformation methods.

Published version
Peer-reviewed

Type

Article

Change log

Authors

Barbrook, Adrian C 
Howe, Christopher J 

Abstract

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.

Description

Keywords

Animals, Anthozoa, Atrazine, Cell Nucleus, Chloramphenicol, Chloroplast Proteins, Chloroplasts, Dinoflagellida, Drug Resistance, Transformation, Genetic

Journal Title

PLoS One

Conference Name

Journal ISSN

1932-6203
1932-6203

Volume Title

14

Publisher

Public Library of Science (PLoS)
Sponsorship
King Abdullah University of Science and Technology (KAUST) (OCRF-2014-CRG3-2216)
King Abdullah University of Science and Technology (KAUST) (unknown)