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 journalArticlepeer-review

191 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 languageEnglish (US)
Pages (from-to)529-534
Number of pages6
JournalNature Methods
Volume11
Issue number5
DOIs
StatePublished - May 2014
Externally publishedYes

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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