Random and targeted transgene insertion in Caenorhabditis elegans using a modified Mos1 transposon

Christian Frøkjær-Jensen; M. Wayne Davis; Mihail Sarov; Jon Taylor; Stephane Flibotte; Matthew LaBella; Andrei Pozniakovsky; Donald G. Moerman; Erik M. Jorgensen

doi:10.1038/nmeth.2889

Random and targeted transgene insertion in Caenorhabditis elegans using a modified Mos1 transposon

Christian Frøkjær-Jensen^*, M. Wayne Davis, Mihail Sarov, Jon Taylor, Stephane Flibotte, Matthew LaBella, Andrei Pozniakovsky, Donald G. Moerman, Erik M. Jorgensen

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

225 Scopus citations

Abstract

We have generated a recombinant Mos1 transposon that can insert up to 45-kb transgenes into the Caenorhabditis elegans genome. The minimal Mos1 transposon (miniMos) is 550 bp long and inserts DNA into the genome at high frequency (~60% of injected animals). Genetic and antibiotic markers can be used for selection, and the transposon is active in C. elegans isolates and Caenorhabditis briggsae. We used the miniMos transposon to generate six universal Mos1-mediated single-copy insertion (mosSCI) landing sites that allow targeted transgene insertion with a single targeting vector into permissive expression sites on all autosomes. We also generated two collections of strains: a set of bright fluorescent insertions that are useful as dominant, genetic balancers and a set of lacO insertions to track genome position.

Original language	English (US)
Pages (from-to)	529-534
Number of pages	6
Journal	Nature Methods
Volume	11
Issue number	5
DOIs	https://doi.org/10.1038/nmeth.2889
State	Published - May 2014
Externally published	Yes

Bibliographical note

Funding Information:
We thank B. Waterston (University of Washington), A. Sapir and P. Sternberg (California Institute of Technology), and the NemaGENETAG consortium for strains; B. Meyer (UC Berkeley) and P. Meister (University of Bern) for validating lacO insertions; the J. Chin (MRC, University of Cambridge), D. Dupuy (University of Bordeaux), B. Lehner (EMBL-CRG, Systems Biology Unit, Barcelona) and G. Seydoux (John Hopkins University) labs for plasmids; M. Maduro (UC Riverside) for improving mosSCI insertion frequency; and K. Hoe for expert technical assistance. Some strains were provided by the Caenorhabditis Genetics Center (CGC), which is funded by US National Institutes of Health (NIH) Office of Research Infrastructure Programs (P40 OD010440). This work was supported by the Carlsberg Foundation (C.F.-J.), NIH grant 1R01GM095817 (E.M.J.), US National Science Foundation grant NSF IOS-0920069 (E.M.J.) and the Howard Hughes Medical Institute (E.M.J.). The Mosmid engineering work was supported by the Max Planck Society (MPG) Initiative “BAC TransgeneOmics” and the NIH ModENCODE project. Work in the laboratory of D.G.M. was supported by the Canadian Institute for Health Research. Work in the laboratory of D.G.M. was supported by the Canadian Institute for Health Research and the Canadian Institute for Advanced Research.

ASJC Scopus subject areas

Biotechnology
Biochemistry
Molecular Biology
Cell Biology

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10.1038/nmeth.2889

Cite this

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title = "Random and targeted transgene insertion in Caenorhabditis elegans using a modified Mos1 transposon",

abstract = "We have generated a recombinant Mos1 transposon that can insert up to 45-kb transgenes into the Caenorhabditis elegans genome. The minimal Mos1 transposon (miniMos) is 550 bp long and inserts DNA into the genome at high frequency (~60% of injected animals). Genetic and antibiotic markers can be used for selection, and the transposon is active in C. elegans isolates and Caenorhabditis briggsae. We used the miniMos transposon to generate six universal Mos1-mediated single-copy insertion (mosSCI) landing sites that allow targeted transgene insertion with a single targeting vector into permissive expression sites on all autosomes. We also generated two collections of strains: a set of bright fluorescent insertions that are useful as dominant, genetic balancers and a set of lacO insertions to track genome position.",

author = "Christian Fr{\o}kj{\ae}r-Jensen and Davis, {M. Wayne} and Mihail Sarov and Jon Taylor and Stephane Flibotte and Matthew LaBella and Andrei Pozniakovsky and Moerman, {Donald G.} and Jorgensen, {Erik M.}",

note = "Funding Information: We thank B. Waterston (University of Washington), A. Sapir and P. Sternberg (California Institute of Technology), and the NemaGENETAG consortium for strains; B. Meyer (UC Berkeley) and P. Meister (University of Bern) for validating lacO insertions; the J. Chin (MRC, University of Cambridge), D. Dupuy (University of Bordeaux), B. Lehner (EMBL-CRG, Systems Biology Unit, Barcelona) and G. Seydoux (John Hopkins University) labs for plasmids; M. Maduro (UC Riverside) for improving mosSCI insertion frequency; and K. Hoe for expert technical assistance. Some strains were provided by the Caenorhabditis Genetics Center (CGC), which is funded by US National Institutes of Health (NIH) Office of Research Infrastructure Programs (P40 OD010440). This work was supported by the Carlsberg Foundation (C.F.-J.), NIH grant 1R01GM095817 (E.M.J.), US National Science Foundation grant NSF IOS-0920069 (E.M.J.) and the Howard Hughes Medical Institute (E.M.J.). The Mosmid engineering work was supported by the Max Planck Society (MPG) Initiative “BAC TransgeneOmics” and the NIH ModENCODE project. Work in the laboratory of D.G.M. was supported by the Canadian Institute for Health Research. Work in the laboratory of D.G.M. was supported by the Canadian Institute for Health Research and the Canadian Institute for Advanced Research.",

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AU - LaBella, Matthew

AU - Pozniakovsky, Andrei

AU - Moerman, Donald G.

AU - Jorgensen, Erik M.

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Random and targeted transgene insertion in Caenorhabditis elegans using a modified Mos1 transposon

Abstract

Bibliographical note

ASJC Scopus subject areas

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