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.
Original language | English |
---|---|
Pages (from-to) | 481-494 |
Number of pages | 14 |
Journal | Nature Methods |
Volume | 17 |
Issue number | 5 |
DOIs | |
Publication status | Published - 2020 May 1 |
Externally published | Yes |
ASJC Scopus subject areas
- Biotechnology
- Biochemistry
- Molecular Biology
- Cell Biology
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Genetic tool development in marine protists : emerging model organisms for experimental cell biology. / Faktorová, Drahomíra; Nisbet, R. Ellen R.; Fernández Robledo, José A.; Casacuberta, Elena; Sudek, Lisa; Allen, Andrew E.; Ares, Manuel; Aresté, Cristina; Balestreri, Cecilia; Barbrook, Adrian C.; Beardslee, Patrick; Bender, Sara; Booth, David S.; Bouget, François Yves; Bowler, Chris; Breglia, Susana A.; Brownlee, Colin; Burger, Gertraud; Cerutti, Heriberto; Cesaroni, Rachele; Chiurillo, Miguel A.; Clemente, Thomas; Coles, Duncan B.; Collier, Jackie L.; Cooney, Elizabeth C.; Coyne, Kathryn; Docampo, Roberto; Dupont, Christopher L.; Edgcomb, Virginia; Einarsson, Elin; Elustondo, Pía A.; Federici, Fernan; Freire-Beneitez, Veronica; Freyria, Nastasia J.; Fukuda, Kodai; García, Paulo A.; Girguis, Peter R.; Gomaa, Fatma; Gornik, Sebastian G.; Guo, Jian; Hampl, Vladimír; Hanawa, Yutaka; Haro-Contreras, Esteban R.; Hehenberger, Elisabeth; Highfield, Andrea; Hirakawa, Yoshihisa; Hopes, Amanda; Howe, Christopher J.; Hu, Ian; Ibañez, Jorge; Irwin, Nicholas A.T.; Ishii, Yuu; Janowicz, Natalia Ewa; Jones, Adam C.; Kachale, Ambar; Fujimura-Kamada, Konomi; Kaur, Binnypreet; Kaye, Jonathan Z.; Kazana, Eleanna; Keeling, Patrick J.; King, Nicole; Klobutcher, Lawrence A.; Lander, Noelia; Lassadi, Imen; Li, Zhuhong; Lin, Senjie; Lozano, Jean Claude; Luan, Fulei; Maruyama, Shinichiro; Matute, Tamara; Miceli, Cristina; Minagawa, Jun; Moosburner, Mark; Najle, Sebastián R.; Nanjappa, Deepak; Nimmo, Isabel C.; Noble, Luke; Novák Vanclová, Anna M.G.; Nowacki, Mariusz; Nuñez, Isaac; Pain, Arnab; Piersanti, Angela; Pucciarelli, Sandra; Pyrih, Jan; Rest, Joshua S.; Rius, Mariana; Robertson, Deborah; Ruaud, Albane; Ruiz-Trillo, Iñaki; Sigg, Monika A.; Silver, Pamela A.; Slamovits, Claudio H.; Jason Smith, G.; Sprecher, Brittany N.; Stern, Rowena; Swart, Estienne C.; Tsaousis, Anastasios D.; Tsypin, Lev; Turkewitz, Aaron; Turnšek, Jernej; Valach, Matus; Vergé, Valérie; von Dassow, Peter; von der Haar, Tobias; Waller, Ross F.; Wang, Lu; Wen, Xiaoxue; Wheeler, Glen; Woods, April; Zhang, Huan; Mock, Thomas; Worden, Alexandra Z.; Lukeš, Julius.
In: Nature Methods, Vol. 17, No. 5, 01.05.2020, p. 481-494.Research output: Contribution to journal › Article › peer-review
}
TY - JOUR
T1 - Genetic tool development in marine protists
T2 - emerging model organisms for experimental cell biology
AU - Faktorová, Drahomíra
AU - Nisbet, R. Ellen R.
AU - Fernández Robledo, José A.
AU - Casacuberta, Elena
AU - Sudek, Lisa
AU - Allen, Andrew E.
AU - Ares, Manuel
AU - Aresté, Cristina
AU - Balestreri, Cecilia
AU - Barbrook, Adrian C.
AU - Beardslee, Patrick
AU - Bender, Sara
AU - Booth, David S.
AU - Bouget, François Yves
AU - Bowler, Chris
AU - Breglia, Susana A.
AU - Brownlee, Colin
AU - Burger, Gertraud
AU - Cerutti, Heriberto
AU - Cesaroni, Rachele
AU - Chiurillo, Miguel A.
AU - Clemente, Thomas
AU - Coles, Duncan B.
AU - Collier, Jackie L.
AU - Cooney, Elizabeth C.
AU - Coyne, Kathryn
AU - Docampo, Roberto
AU - Dupont, Christopher L.
AU - Edgcomb, Virginia
AU - Einarsson, Elin
AU - Elustondo, Pía A.
AU - Federici, Fernan
AU - Freire-Beneitez, Veronica
AU - Freyria, Nastasia J.
AU - Fukuda, Kodai
AU - García, Paulo A.
AU - Girguis, Peter R.
AU - Gomaa, Fatma
AU - Gornik, Sebastian G.
AU - Guo, Jian
AU - Hampl, Vladimír
AU - Hanawa, Yutaka
AU - Haro-Contreras, Esteban R.
AU - Hehenberger, Elisabeth
AU - Highfield, Andrea
AU - Hirakawa, Yoshihisa
AU - Hopes, Amanda
AU - Howe, Christopher J.
AU - Hu, Ian
AU - Ibañez, Jorge
AU - Irwin, Nicholas A.T.
AU - Ishii, Yuu
AU - Janowicz, Natalia Ewa
AU - Jones, Adam C.
AU - Kachale, Ambar
AU - Fujimura-Kamada, Konomi
AU - Kaur, Binnypreet
AU - Kaye, Jonathan Z.
AU - Kazana, Eleanna
AU - Keeling, Patrick J.
AU - King, Nicole
AU - Klobutcher, Lawrence A.
AU - Lander, Noelia
AU - Lassadi, Imen
AU - Li, Zhuhong
AU - Lin, Senjie
AU - Lozano, Jean Claude
AU - Luan, Fulei
AU - Maruyama, Shinichiro
AU - Matute, Tamara
AU - Miceli, Cristina
AU - Minagawa, Jun
AU - Moosburner, Mark
AU - Najle, Sebastián R.
AU - Nanjappa, Deepak
AU - Nimmo, Isabel C.
AU - Noble, Luke
AU - Novák Vanclová, Anna M.G.
AU - Nowacki, Mariusz
AU - Nuñez, Isaac
AU - Pain, Arnab
AU - Piersanti, Angela
AU - Pucciarelli, Sandra
AU - Pyrih, Jan
AU - Rest, Joshua S.
AU - Rius, Mariana
AU - Robertson, Deborah
AU - Ruaud, Albane
AU - Ruiz-Trillo, Iñaki
AU - Sigg, Monika A.
AU - Silver, Pamela A.
AU - Slamovits, Claudio H.
AU - Jason Smith, G.
AU - Sprecher, Brittany N.
AU - Stern, Rowena
AU - Swart, Estienne C.
AU - Tsaousis, Anastasios D.
AU - Tsypin, Lev
AU - Turkewitz, Aaron
AU - Turnšek, Jernej
AU - Valach, Matus
AU - Vergé, Valérie
AU - von Dassow, Peter
AU - von der Haar, Tobias
AU - Waller, Ross F.
AU - Wang, Lu
AU - Wen, Xiaoxue
AU - Wheeler, Glen
AU - Woods, April
AU - Zhang, Huan
AU - Mock, Thomas
AU - Worden, Alexandra Z.
AU - Lukeš, Julius
N1 - Funding Information: GBMF4962 to J.A.F.R.; GBMF4980 and 4980.01 to S.L.; GBMF 4977 and 4977.01 to R.F.W.; GBMF4962.01 to C.H.S.; GBMF4985 to J.M.; GBMF4976 and 4976.01 to C.H.; GBMF4963 and 4963.01 to V.E.; GBMF5007 to C.L.D.; GBMF4983 and 4983.01 to J.L.; GBMF4975 and 4975.01 to A.D.T.; GBMF4973 and 4973.01 to I.R.-T. and GBMF4965 to N.K.), by The Leverhulme Trust (RPG-2017-364) to T. Mock and A. Hopes, and by ERD funds (16_019/0000759) from the Czech Ministry of Education to J.L. Funding Information: We thank M. Salisbury and D. Lacono, C. Poirier, M. Hamilton, C. Eckmann, H. Igel, C. Yung and K. Hoadley for assistance; V.K. Nagarajan, M. Accerbi and P.J. Green who carried out Agrobacterium studies in Heterosigma akashiwo, and N. Kraeva, C. Bianchi and V. Yurchenko for the help with designing the p57-V5+NeoR construct. We are also grateful to the protocols.io team (L. Teytelman and A. Broellochs) for their support. This collaborative effort was supported by the Gordon and Betty Moore Foundation EMS Program of the Marine Microbiology Initiative (grant nos. GBMF4972 and 4972.01 to F.-Y.B.; GBMF4970 and 4970.01 to M.A. and A.Z.W.; GBMF3788 to A.Z.W.; GBMF 4968 and 4968.01 to H.C.; GBMF4984 to V.H.; GBMF4974 and 4974.01 to C. Brownlee; GBMF4964 to Y. Hirakawa; GBMF4961 to T. Mock; GBMF4958 to P.S.; GBMF4957 to A.T.; GBMF4960 to G.J.S.; GBMF4979 to K.C.; GBMF4982 and 4982.01 to J.L.C.; GBMF4964 to P.J.K.; GBMF4981 to P.v.D.; GBMF5006 to A.E.A.; GBMF4986 to C.M.; Publisher Copyright: © 2020, The Author(s), under exclusive licence to Springer Nature America, Inc.
PY - 2020/5/1
Y1 - 2020/5/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85083247242&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85083247242&partnerID=8YFLogxK
U2 - 10.1038/s41592-020-0796-x
DO - 10.1038/s41592-020-0796-x
M3 - Article
C2 - 32251396
AN - SCOPUS:85083247242
VL - 17
SP - 481
EP - 494
JO - Nature Methods
JF - Nature Methods
SN - 1548-7091
IS - 5
ER -