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Genetic tool development in marine protists: Emerging model organisms for experimental cell biology

Drahomíra, Faktorová and Tsypin, Lev (2020) Genetic tool development in marine protists: Emerging model organisms for experimental cell biology. Nature Methods, 17 (5). pp. 481-494. ISSN 1548-7091. PMCID PMC7200600; PMC7200595. https://resolver.caltech.edu/CaltechAUTHORS:20190802-133147287

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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.


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https://doi.org/10.1038/s41592-020-0796-xDOIArticle
https://rdcu.be/b3taPPublisherFree ReadCube access
https://doi.org/10.1038/s41592-020-0828-6DOIPublisher Correction
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https://doi.org/10.1101/718239DOIDiscussion Paper
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7200600/PubMed CentralArticle
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7200595/PubMed CentralPublisher Correction
ORCID:
AuthorORCID
Drahomíra, Faktorová0000-0001-9623-2233
Tsypin, Lev0000-0002-0642-8468
Additional Information:© 2020 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Received 26 June 2019; Accepted 02 March 2020; Published 06 April 2020. 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.; 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. Data availability: The data that support the findings of this study are available from the corresponding authors as well as the other authors upon request (for the contacts see Supplementary Table 6). Source data for Figs. 3 and 4 and Supplementary Figs. 9b,c, 11a and 12b,c are available online. Author Contributions: The project was conceived and designed by A.C.J., J.Z.K., S.B., D.F., J.L., R.E.R.N., J.A.F.R., E.C., L.S., A.Z.W., T. Mock, A.E.A., F.-Y.B, C. Brownlee, C. Bowler, H.C., T.C., J.L.C., K.C., C.L.D., V.E., V.H., Y. Hirakawa, C.J.H., P.J.K., N.K., S.L., C.M., J.M., I.R.-T., P.A.S., C.H.S., G.J.S., A.D.T., P.v.D., A.T. and R.F.W. Data analysis was carried out by M.A.Jr, C.A., C. Balestreri, A.C.B., P.B., D.S.B., S.A.B., G.B., R.C., M.A.C., D.B.C., E.C.C., R.D., E.E., P.A.E., F.F., V.F.-B., N.J.F., K.F., P.A.G., P.R.G., F.G., S.G.G., J.G., Y. Hanawa, E.R.H.-C., E.H., A. Highfield, A. Hopes, I.H., J.I., N.A.T.I., Y.I., N.E.J., A.K., K.F.-K., B.K., E.K., L.A.K., N.L., I.L., Z.L., J.-C.L., F.L., S.M., T. Matute, M.M., S.R.N., D.N., I.C.N., L.N., A.M.G.N.V., M.N., I.N., A. Pain, A. Piersanti, S.P., J.P., J.S.R., M.R., D.R., A.R., M.A.S., E.C.S., B.N.S., R.S., T.v.d.H., L.T., J.T., M.V., V.V., L.W., X.W., G.W., A.W. and H.Z. The manuscript was written by D.F., R.E.R.N., J.A.F.R., E.C., L.S., T. Mock, A.Z.W. and J.L. with input from all authors. The authors declare no competing interests.
Errata:This paper was originally published under standard Springer Nature copyright (© The Author(s), under exclusive licence to Springer Nature America, Inc.). It is now available as an open-access paper under a Creative Commons Attribution 4.0 International license. The error has been corrected in the print, PDF and HTML versions of the article.
Funders:
Funding AgencyGrant Number
Gordon and Betty Moore FoundationGBMF4972
Gordon and Betty Moore Foundation4972.01
Gordon and Betty Moore FoundationGBMF4970
Gordon and Betty Moore Foundation4970.01
Gordon and Betty Moore FoundationGBMF3788
Gordon and Betty Moore FoundationGBMF 4968
Gordon and Betty Moore Foundation4968.01
Gordon and Betty Moore FoundationGBMF4984
Gordon and Betty Moore FoundationGBMF4974
Gordon and Betty Moore Foundation4974.01
Gordon and Betty Moore FoundationGBMF4964
Gordon and Betty Moore FoundationGBMF4961
Gordon and Betty Moore FoundationGBMF4958
Gordon and Betty Moore FoundationGBMF4957
Gordon and Betty Moore FoundationGBMF4960
Gordon and Betty Moore FoundationGBMF4979
Gordon and Betty Moore FoundationGBMF4982
Gordon and Betty Moore Foundation4982.01
Gordon and Betty Moore FoundationGBMF4964
Gordon and Betty Moore FoundationGBMF4981
Gordon and Betty Moore FoundationGBMF5006
Gordon and Betty Moore FoundationGBMF4986
Gordon and Betty Moore FoundationGBMF4962
Gordon and Betty Moore FoundationGBMF4980
Gordon and Betty Moore Foundation4980.01
Gordon and Betty Moore FoundationGBMF 4977
Gordon and Betty Moore Foundation4977.01
Gordon and Betty Moore FoundationGBMF4962.01
Gordon and Betty Moore FoundationGBMF4985
Gordon and Betty Moore FoundationGBMF4976
Gordon and Betty Moore Foundation4976.01
Gordon and Betty Moore FoundationGBMF4963
Gordon and Betty Moore Foundation4963.01
Gordon and Betty Moore FoundationGBMF5007
Gordon and Betty Moore FoundationGBMF4983
Gordon and Betty Moore Foundation4983.01
Gordon and Betty Moore FoundationGBMF4975
Gordon and Betty Moore Foundation4975.01
Gordon and Betty Moore FoundationGBMF4973
Gordon and Betty Moore Foundation4973.01
Gordon and Betty Moore FoundationGBMF4965
Leverhulme TrustRPG-2017-364
European Regional Development Fund16_019/0000759
Czech Ministry of EducationUNSPECIFIED
Subject Keywords:Genetic models; Molecular biology
Issue or Number:5
PubMed Central ID:PMC7200600; PMC7200595
Record Number:CaltechAUTHORS:20190802-133147287
Persistent URL:https://resolver.caltech.edu/CaltechAUTHORS:20190802-133147287
Official Citation:Faktorová, D., Nisbet, R.E.R., Fernández Robledo, J.A. et al. Genetic tool development in marine protists: emerging model organisms for experimental cell biology. Nat Methods 17, 481–494 (2020). https://doi.org/10.1038/s41592-020-0796-x
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:97621
Collection:CaltechAUTHORS
Deposited By: George Porter
Deposited On:05 Aug 2019 14:57
Last Modified:24 Nov 2020 19:12

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