Nat Methods. 2020 May;17(5):481-494. doi: 10.1038/s41592-020-0796-x. | PubMed

Drahomíra Faktorová¹, R Ellen R Nisbet^2,3, José A Fernández Robledo⁴, Elena Casacuberta⁵, Lisa Sudek⁶, Andrew E Allen^7,8, Manuel Ares Jr⁹, Cristina Aresté⁵, Cecilia Balestreri¹⁰, Adrian C Barbrook², Patrick Beardslee¹¹, Sara Bender¹², David S Booth¹³, François-Yves Bouget¹⁴, Chris Bowler¹⁵, Susana A Breglia¹⁶, Colin Brownlee¹⁰, Gertraud Burger¹⁷, Heriberto Cerutti¹¹, Rachele Cesaroni¹⁸, Miguel A Chiurillo¹⁹, Thomas Clemente¹¹, Duncan B Coles⁴, Jackie L Collier²⁰, Elizabeth C Cooney²¹, Kathryn Coyne²², Roberto Docampo¹⁹, Christopher L Dupont⁸, Virginia Edgcomb²³, Elin Einarsson², Pía A Elustondo^16,24, Fernan Federici²⁵, Veronica Freire-Beneitez^26,27, Nastasia J Freyria⁴, Kodai Fukuda²⁸, Paulo A García²⁹, Peter R Girguis³⁰, Fatma Gomaa³⁰, Sebastian G Gornik³¹, Jian Guo^6,9, Vladimír Hampl³², Yutaka Hanawa³³, Esteban R Haro-Contreras¹⁶, Elisabeth Hehenberger²¹, Andrea Highfield¹⁰, Yoshihisa Hirakawa³³, Amanda Hopes³⁴, Christopher J Howe², Ian Hu², Jorge Ibañez²⁵, Nicholas A T Irwin²¹, Yuu Ishii³⁵, Natalia Ewa Janowicz³², Adam C Jones¹², Ambar Kachale³⁶, Konomi Fujimura-Kamada³⁷, Binnypreet Kaur³⁶, Jonathan Z Kaye¹², Eleanna Kazana^26,27, Patrick J Keeling²¹, Nicole King¹³, Lawrence A Klobutcher³⁸, Noelia Lander¹⁹, Imen Lassadi², Zhuhong Li¹⁹, Senjie Lin³⁸, Jean-Claude Lozano¹⁴, Fulei Luan¹¹, Shinichiro Maruyama³⁵, Tamara Matute²⁵, Cristina Miceli³⁹, Jun Minagawa^37,40, Mark Moosburner^7,8, Sebastián R Najle^5,41, Deepak Nanjappa²², Isabel C Nimmo², Luke Noble^42,43, Anna M G Novák Vanclová³², Mariusz Nowacki¹⁸, Isaac Nuñez²⁵, Arnab Pain^44,45, Angela Piersanti³⁹, Sandra Pucciarelli³⁹, Jan Pyrih^26,32, Joshua S Rest⁴⁶, Mariana Rius²⁰, Deborah Robertson⁴⁷, Albane Ruaud^25,48, Iñaki Ruiz-Trillo^5,49,50, Monika A Sigg¹³, Pamela A Silver^51,52, Claudio H Slamovits¹⁶, G Jason Smith⁵³, Brittany N Sprecher³⁸, Rowena Stern¹⁰, Estienne C Swart^18,48, Anastasios D Tsaousis^26,27, Lev Tsypin^54,55, Aaron Turkewitz⁵⁴, Jernej Turnšek^7,8,51,52, Matus Valach¹⁷, Valérie Vergé¹⁴, Peter von Dassow^25,56, Tobias von der Haar²⁶, Ross F Waller², Lu Wang⁵⁷, Xiaoxue Wen¹¹, Glen Wheeler¹⁰, April Woods⁵³, Huan Zhang³⁸, Thomas Mock⁵⁸, Alexandra Z Worden^59,60, Julius Lukeš⁶¹

1. Institute of Parasitology, Biology Centre, Czech Academy of Sciences and Faculty of Sciences, University of South Bohemia, České Budějovice, Czech Republic. dranov@paru.cas.cz.
2. Department of Biochemistry, University of Cambridge, Cambridge, UK.
3. School of Biosciences, University of Nottingham, Sutton Bonington, UK.
4. Bigelow Laboratory for Ocean Sciences, East Boothbay, ME, USA.
5. Institut de Biologia Evolutiva, CSIC-Universitat Pompeu Fabra, Barcelona, Spain.
6. Monterey Bay Aquarium Research Institute, Moss Landing, CA, USA.
7. Integrative Oceanography Division, Scripps Institution of Oceanography, University of California, San Diego, CA, USA.
8. Microbial and Environmental Genomics, J. Craig Venter Institute, La Jolla, CA, USA.
9. Molecular, Cell and Developmental Biology, University of California, Santa Cruz, CA, USA.
10. The Marine Biological Association, Plymouth and School of Ocean and Earth Sciences, University of Southampton, Southampton, UK.
11. School of Biological Sciences, University of Nebraska, Lincoln, NE, USA.
12. Gordon and Betty Moore Foundation, Palo Alto, CA, USA.
13. Department of Molecular and Cell Biology, University of California, Berkeley, CA, USA.
14. Sorbonne Université, CNRS UMR7621, Observatoire Océanologique, Banyuls sur Mer, France.
15. Institut de Biologie de l'Ecole Normale Supérieure (IBENS), Ecole Normale Supérieure, CNRS, INSERM, Université PSL, Paris, France.
16. Centre for Comparative Genomics and Evolutionary Bioinformatics, Dalhousie University, Halifax, Nova Scotia, Canada.
17. Department of Biochemistry and Robert-Cedergren Centre for Bioinformatics and Genomics, Université de Montréal, Montreal, Quebec, Canada.
18. Institute of Cell Biology, University of Bern, Bern, Switzerland.
19. Center for Tropical and Emerging Global Diseases, University of Georgia, Athens, GA, USA.
20. School of Marine and Atmospheric Sciences, Stony Brook University, Stony Brook, NY, USA.
21. Department of Botany, University of British Columbia, Vancouver, British Columbia, Canada.
22. University of Delaware College of Earth, Ocean and Environment, Lewes, DE, USA.
23. Woods Hole Oceanographic Institution, Woods Hole, MA, USA.
24. AGADA Biosciences Inc., Halifax, Nova Scotia, Canada.
25. Facultad Ciencias Biológicas, Pontificia Universidad Católica de Chile, Fondo de Desarrollo de Areas Prioritarias, Center for Genome Regulation and Millennium Institute for Integrative Biology (iBio), Santiago de Chile, Chile.
26. School of Biosciences, University of Kent, Canterbury, Kent, UK.
27. Laboratory of Molecular and Evolutionary Parasitology, University of Kent, Kent, UK.
28. Graduate School of Life and Environmental Sciences, University of Tsukuba, Ibaraki, Japan.
29. Department of Mechanical Engineering, Massachusetts Institute of Technology, Boston, MA, USA.
30. Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, MA, USA.
31. Centre for Organismal Studies, University of Heidelberg, Heidelberg, Germany.
32. Department of Parasitology, Faculty of Science, Charles University, BIOCEV, Vestec, Czech Republic.
33. Faculty of Life and Environmental Sciences, University of Tsukuba, Ibaraki, Japan.
34. School of Environmental Sciences, University of East Anglia, Norwich, UK.
35. Graduate School of Life Sciences, Tohoku University, Sendai, Miyagi, Japan.
36. Institute of Parasitology, Biology Centre, Czech Academy of Sciences and Faculty of Sciences, University of South Bohemia, České Budějovice, Czech Republic.
37. Division of Environmental Photobiology, National Institute for Basic Biology, Okazaki, Aichi, Japan.
38. Department of Marine Sciences, University of Connecticut , Groton, CT, USA.
39. School of Biosciences and Veterinary Medicine, University of Camerino, Camerino, Italy.
40. Department of Basic Biology, School of Life Science, Graduate University for Advanced Studies, Okazaki, Aichi, Japan.
41. Instituto de Biología Molecular y Celular, CONICET, and Facultad de Ciencias Bioquímicas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina.
42. Center for Genomics and Systems Biology, New York University, New York, NY, USA.
43. Institute de Biologie de l'ENS, Département de biologie, École Normale Supérieure, CNRS, INSERM, Paris, France.
44. Biological and Environmental Science and Engineering Division, King Abdullah University of Science and Technology, Thuwal, Saudi Arabia.
45. Center for Zoonosis Control, Global Institution for Collaborative Research and Education, Hokkaido University, Sapporo, Japan.
46. Department of Ecology and Evolution, Stony Brook University, Stony Brook, NY, USA.
47. Lasry Center for Biosciences, Clark University, Worcester, MA, USA.
48. Max Planck Institute for Developmental Biology, Tübingen, Germany.
49. Departament de Genètica Microbiologia i Estadıśtica, Universitat de Barcelona, Barcelona, Spain.
50. Catalan Institution for Research and Advanced Studies, Barcelona, Spain.
51. Department of Systems Biology, Harvard Medical School, Boston, MA, USA.
52. Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA, USA.
53. Department of Environmental Biotechnology, Moss Landing Marine Laboratories, Moss Landing, CA, USA.
54. Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, IL, USA.
55. Department of Biology, California Institute of Technology, Pasadena, CA, USA.
56. Instituto Milenio de Oceanografia de Chile, Concepción, Chile.
57. Institute of Oceanography, Minjiang University, Fuzhou, China.
58. School of Environmental Sciences, University of East Anglia, Norwich, UK. t.mock@uea.ac.uk.
59. Monterey Bay Aquarium Research Institute, Moss Landing, CA, USA. azworden@geomar.de.
60. Ocean EcoSystems Biology Unit, Marine Ecology Division, Helmholtz Centre for Ocean Research, Kiel, Germany. azworden@geomar.de.
61. Institute of Parasitology, Biology Centre, Czech Academy of Sciences and Faculty of Sciences, University of South Bohemia, České Budějovice, Czech Republic. jula@paru.cas.cz.

Abstract

Diverse microbial ecosystems underpin life in the sea. Among these microbes are many unicellular eukaryotes that span the diversity of the eukaryotic tree of life. However, genetic tractability has been limited to a few species, which do not represent eukaryotic diversity or environmentally relevant taxa. Here, we report on the development of genetic tools in a range of protists primarily from marine environments. We present evidence for foreign DNA delivery and expression in 13 species never before transformed and for advancement of tools for eight other species, as well as potential reasons for why transformation of yet another 17 species tested was not achieved. Our resource in genetic manipulation will provide insights into the ancestral eukaryotic lifeforms, general eukaryote cell biology, protein diversification and the evolution of cellular pathways.

Presented By Drahomíra Faktorová | ORCID iD