Designer diatom episomes delivered by bacterial conjugation

Karas, Bogumil J.; Diner, Rachel E.; Lefebvre, Stephane C.; McQuaid, Jeff; Phillips, Alex P. R.; Noddings, Chari M.; Brunson, John K.; Valas, Ruben E.; Deerinck, Thomas J.; Jablanovic, Jelena; Gillard, Jeroen T. F.; Beeri, Karen; Ellisman, Mark H.; Glass, John I.; Hutchison, Clyde A.; Smith, Hamilton O.; Venter, J. Craig; Allen, Andrew E.; Dupont, Christopher L.; Weyman, Philip D.

doi:10.1038/ncomms7925

Designer diatom episomes delivered by bacterial conjugation

Journal Article · Tue Apr 21 00:00:00 EDT 2015 · Nature Communications

DOI:https://doi.org/10.1038/ncomms7925· OSTI ID:1455147

Karas, Bogumil J. ^[1]; Diner, Rachel E. ^[2]; Lefebvre, Stephane C. ^[3]; McQuaid, Jeff ^[3]; Phillips, Alex P. R. ^[4]; Noddings, Chari M. ^[4]; Brunson, John K. ^[4]; Valas, Ruben E. ^[3]; Deerinck, Thomas J. ^[5]; Jablanovic, Jelena ^[3]; Gillard, Jeroen T. F. ^[3]; Beeri, Karen ^[3]; Ellisman, Mark H. ^[5]; Glass, John I. ^[4]; Hutchison, Clyde A. ^[4]; Smith, Hamilton O. ^[4]; Venter, J. Craig ^[6]; Allen, Andrew E. ^[2]; Dupont, Christopher L. ^[3]; Weyman, Philip D. ^[4]

J. Craig Venter Inst., La Jolla, CA (United States). Synthetic Biology and Bioenergy Group; Synthetic Biology and Bioenergy Group, J. Craig Venter Institute, La Jolla, California 92037
J. Craig Venter Inst., La Jolla, CA (United States). Microbial and Environmental Genomics Group; Univ. of California, San Diego, CA (United States). Integrative Oceanography Division. Scripps Inst. of Oceanography
J. Craig Venter Inst., La Jolla, CA (United States). Microbial and Environmental Genomics Group
J. Craig Venter Inst., La Jolla, CA (United States). Synthetic Biology and Bioenergy Group
Univ. of California, San Diego, CA (United States). National Center for Microscopy and Imaging Research
J. Craig Venter Inst., La Jolla, CA (United States). Synthetic Biology and Bioenergy Group. Microbial and Environmental Genomics Group

Eukaryotic microalgae hold great promise for the bioproduction of fuels and higher value chemicals. However, compared with model genetic organisms such as Escherichia coli and Saccharomyces cerevisiae, characterization of the complex biology and biochemistry of algae and strain improvement has been hampered by the inefficient genetic tools. To date, many algal species are transformable only via particle bombardment, and the introduced DNA is integrated randomly into the nuclear genome. Here we describe the first nuclear episomal vector for diatoms and a plasmid delivery method via conjugation from Escherichia coli to the diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana. We identify a yeast-derived sequence that enables stable episome replication in these diatoms even in the absence of antibiotic selection and show that episomes are maintained as closed circles at copy number equivalent to native chromosomes. This highly efficient genetic system facilitates high-throughput functional characterization of algal genes and accelerates molecular phytoplankton research.

View Accepted Manuscript (DOE)

Research Organization:: J. Craig Venter Inst., La Jolla, CA (United States)

Sponsoring Organization:: Gordon and Betty Moore Foundation (United States); Natural Sciences and Engineering Research Council of Canada (NSERC); Synthetic Genomics Inc. (United States); USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)

Grant/Contract Number:: SC0008593

OSTI ID:: 1455147

Journal Information:: Nature Communications, Journal Name: Nature Communications Vol. 6; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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Terpenoid Metabolic Engineering in Photosynthetic Microorganisms Vavitsas, Konstantinos; Fabris, Michele; Vickers, Claudia Genes, Vol. 9, Issue 11 https://doi.org/10.3390/genes9110520	journal	October 2018
Proximity proteomics in a marine diatom reveals a putative cell surface-to-chloroplast iron trafficking pathway Turnšek, Jernej; Brunson, John K.; Viedma, Maria del Pilar Martinez eLife, Vol. 10 https://doi.org/10.7554/elife.52770	journal	February 2021
Assessment of genomic changes in a CRISPR/Cas9 Phaeodactylum tricornutum mutant through whole genome resequencing Russo, Monia Teresa; Aiese Cigliano, Riccardo; Sanseverino, Walter PeerJ, Vol. 6 https://doi.org/10.7717/peerj.5507	journal	January 2018
Blasticidin-S deaminase, a new selection marker for genetic transformation of the diatom Phaeodactylum tricornutum Buck, Jochen M.; Río Bártulos, Carolina; Gruber, Ansgar PeerJ, Vol. 6 https://doi.org/10.7717/peerj.5884	journal	January 2018

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