Single-copy insertion of transgenes in Caenorhabditis elegans.
Research output: Contribution to journal › Journal article › Research › peer-review
Standard
Single-copy insertion of transgenes in Caenorhabditis elegans. / Frøkjaer-Jensen, Christian; Davis, M Wayne; Hopkins, Christopher E; Newman, Blake J; Thummel, Jason M; Olesen, Søren-Peter; Grunnet, Morten; Jorgensen, Erik M.
In: Nature Genetics, Vol. 40, No. 11, 2008, p. 1375-83.Research output: Contribution to journal › Journal article › Research › peer-review
Harvard
APA
Vancouver
Author
Bibtex
}
RIS
TY - JOUR
T1 - Single-copy insertion of transgenes in Caenorhabditis elegans.
AU - Frøkjaer-Jensen, Christian
AU - Davis, M Wayne
AU - Hopkins, Christopher E
AU - Newman, Blake J
AU - Thummel, Jason M
AU - Olesen, Søren-Peter
AU - Grunnet, Morten
AU - Jorgensen, Erik M
PY - 2008
Y1 - 2008
N2 - 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.
AB - 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.
U2 - 10.1038/ng.248
DO - 10.1038/ng.248
M3 - Journal article
C2 - 18953339
VL - 40
SP - 1375
EP - 1383
JO - Nature Genetics
JF - Nature Genetics
SN - 1061-4036
IS - 11
ER -
ID: 8418416