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.

AGADA Biosciences Inc Halifax Nova Scotia Canada

Bigelow Laboratory for Ocean Sciences East Boothbay ME USA

Biological and Environmental Science and Engineering Division King Abdullah University of Science and Technology Thuwal Saudi Arabia

Catalan Institution for Research and Advanced Studies Barcelona Spain

Center for Genomics and Systems Biology New York University New York NY USA

Center for Tropical and Emerging Global Diseases University of Georgia Athens GA USA

Center for Zoonosis Control Global Institution for Collaborative Research and Education Hokkaido University Sapporo Japan

Centre for Comparative Genomics and Evolutionary Bioinformatics Dalhousie University Halifax Nova Scotia Canada

Centre for Organismal Studies University of Heidelberg Heidelberg Germany

Departament de Genètica Microbiologia i Estadıśtica Universitat de Barcelona Barcelona Spain

Department of Basic Biology School of Life Science Graduate University for Advanced Studies Okazaki Aichi Japan

Department of Biochemistry and Robert Cedergren Centre for Bioinformatics and Genomics Université de Montréal Montreal Quebec Canada

Department of Biochemistry University of Cambridge Cambridge UK

Department of Biology California Institute of Technology Pasadena CA USA

Department of Botany University of British Columbia Vancouver British Columbia Canada

Department of Ecology and Evolution Stony Brook University Stony Brook NY USA

Department of Environmental Biotechnology Moss Landing Marine Laboratories Moss Landing CA USA

Department of Marine Sciences University of Connecticut Groton CT USA

Department of Mechanical Engineering Massachusetts Institute of Technology Boston MA USA

Department of Molecular and Cell Biology University of California Berkeley CA USA

Department of Molecular Genetics and Cell Biology University of Chicago Chicago IL USA

Department of Organismic and Evolutionary Biology Harvard University Cambridge MA USA

Department of Parasitology Faculty of Science Charles University BIOCEV Vestec Czech Republic

Department of Systems Biology Harvard Medical School Boston MA USA

Division of Environmental Photobiology National Institute for Basic Biology Okazaki Aichi Japan

Facultad Ciencias Biológicas Pontificia Universidad Católica de Chile Fondo de Desarrollo de Areas Prioritarias Center for Genome Regulation and Millennium Institute for Integrative Biology Santiago de Chile Chile

Faculty of Life and Environmental Sciences University of Tsukuba Ibaraki Japan

Gordon and Betty Moore Foundation Palo Alto CA USA

Graduate School of Life and Environmental Sciences University of Tsukuba Ibaraki Japan

Graduate School of Life Sciences Tohoku University Sendai Miyagi Japan

Institut de Biologia Evolutiva CSIC Universitat Pompeu Fabra Barcelona Spain

Institut de Biologie de l'Ecole Normale Supérieure Ecole Normale Supérieure CNRS INSERM Université PSL Paris France

Institute de Biologie de l'ENS Département de biologie École Normale Supérieure CNRS INSERM Paris France

Institute of Cell Biology University of Bern Bern Switzerland

Institute of Oceanography Minjiang University Fuzhou China

Institute of Parasitology Biology Centre Czech Academy of Sciences and Faculty of Sciences University of South Bohemia České Budějovice Czech Republic

Instituto de Biología Molecular y Celular CONICET and Facultad de Ciencias Bioquímicas y Farmacéuticas Universidad Nacional de Rosario Rosario Argentina

Instituto Milenio de Oceanografia de Chile Concepción Chile

Integrative Oceanography Division Scripps Institution of Oceanography University of California San Diego CA USA

Laboratory of Molecular and Evolutionary Parasitology University of Kent Kent UK

Lasry Center for Biosciences Clark University Worcester MA USA

Max Planck Institute for Developmental Biology Tübingen Germany

Microbial and Environmental Genomics J Craig Venter Institute La Jolla CA USA

Molecular Cell and Developmental Biology University of California Santa Cruz CA USA

Monterey Bay Aquarium Research Institute Moss Landing CA USA

Ocean EcoSystems Biology Unit Marine Ecology Division Helmholtz Centre for Ocean Research Kiel Germany

School of Biological Sciences University of Nebraska Lincoln NE USA

School of Biosciences and Veterinary Medicine University of Camerino Camerino Italy

School of Biosciences University of Kent Canterbury Kent UK

School of Biosciences University of Nottingham Sutton Bonington UK

School of Environmental Sciences University of East Anglia Norwich UK

School of Marine and Atmospheric Sciences Stony Brook University Stony Brook NY USA

Sorbonne Université CNRS UMR7621 Observatoire Océanologique Banyuls sur Mer France

The Marine Biological Association Plymouth and School of Ocean and Earth Sciences University of Southampton Southampton UK

University of Delaware College of Earth Ocean and Environment Lewes DE USA

Woods Hole Oceanographic Institution Woods Hole MA USA

Wyss Institute for Biologically Inspired Engineering Harvard University Boston MA USA

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Nat Methods. 2020 May;17(5):469-470. doi: 10.1038/s41592-020-0794-z. PubMed

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Nat Methods. 2020 May;17(5):551. doi: 10.1038/s41592-020-0828-6. PubMed

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