Single-copy insertion of transgenes in Caenorhabditis elegans

Christian Frøkjær-Jensen, M. Wayne Davis, Christopher E. Hopkins, Blake J. Newman, Jason M. Thummel, Søren Peter Olesen, Morten Grunnet, Erik M. Jorgensen

Research output: Contribution to journalArticlepeer-review

604 Scopus citations

Abstract

At present, transgenes in Caenorhabditis elegans are generated by injecting DNA into the germline. The DNA assembles into a semistable extrachromosomal array composed of many copies of injected DNA. These transgenes are typically overexpressed in somatic cells and silenced in the germline. We have developed a method that inserts a single copy of a transgene into a defined site. Mobilization of a Mos1 transposon generates a double-strand break in noncoding DNA. The break is repaired by copying DNA from an extrachromosomal template into the chromosomal site. Homozygous single-copy insertions can be obtained in less than 2 weeks by injecting approximately 20 worms. We have successfully inserted transgenes as long as 9 kb and verified that single copies are inserted at the targeted site. Single-copy transgenes are expressed at endogenous levels and can be expressed in the female and male germlines.

Original languageEnglish (US)
Pages (from-to)1375-1383
Number of pages9
JournalNature Genetics
Volume40
Issue number11
DOIs
StatePublished - Nov 1 2008
Externally publishedYes

ASJC Scopus subject areas

  • Genetics

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