We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Single-copy insertion of transgenes in Caenorhabditis elegans.
- Authors
Frøkjær-Jensen, Christian; Wayne Davis, M; Hopkins, Christopher E; Newman, Blake J; Thummel, Jason M; Olesen, Søren-Peter; Grunnet, Morten; Jorgensen, Erik M
- 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.
- Publication
Nature Genetics, 2008, Vol 40, Issue 11, p1375
- ISSN
1061-4036
- Publication type
Academic Journal
- DOI
10.1038/ng.248