The Coral Symbiomics Lab
Red Sea Research Center
Division of Biological and Environmental Science and Engineering

JEChen_2019_PlosONE

The Genetic Intractability of Symbiodinium Microadriaticum to Standard Algal Transformation Methods

Chen, Jit Ern, Adrian C. Barbrook, Guoxin Cui, Christopher J. Howe, and Manuel Aranda. “The Genetic Intractability of Symbiodinium Microadriaticum to Standard Algal Transformation Methods.” PLOS ONE 14, no. 2 (February 19, 2019): e0211936. https://doi.org/10.1371/journal.pone.0211936.
Chen Jit Ern, Adrian C. Barbrook, Guoxin Cui, Christopher J. Howe, and Manuel Aranda
Symbiodinium, Transformation methods, CRISPR/cas9
2019
​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.