TY - JOUR
T1 - The genetic intractability of Symbiodinium microadriaticum to standard algal transformation methods
AU - Chen, Jit Ern
AU - Barbrook, Adrian C.
AU - Cui, Guoxin
AU - Howe, Christopher J.
AU - Aranda, Manuel
N1 - KAUST Repository Item: Exported on 2020-10-01
Acknowledged KAUST grant number(s): URF/1/1705-01, OCRF-2014-CRG3-2216
Acknowledgements: Funded by King Abdullah University of Science and Technology (KAUST) Office of Sponsored Research (OSR). https://www.kaust.edu.sa/en/research/sponsored-research URF/1/1705-01. MA. King Abdullah University of Science and Technology (KAUST) Office of Sponsored Research. (OSR). https://www.kaust.edu.sa/en/research/sponsored-research OCRF-2014-CRG3-2216. MA, CJH. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
PY - 2019/2/19
Y1 - 2019/2/19
N2 - 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.
AB - 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.
UR - http://hdl.handle.net/10754/625875
UR - https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0211936
UR - http://www.scopus.com/inward/record.url?scp=85061865891&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0211936
DO - 10.1371/journal.pone.0211936
M3 - Article
C2 - 30779749
SN - 1932-6203
VL - 14
SP - e0211936
JO - PLOS ONE
JF - PLOS ONE
IS - 2
ER -