Single-copy insertion of transgenes in Caenorhabditis elegans.

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

doi:10.1038/ng.248

Single-copy insertion of transgenes in Caenorhabditis elegans.

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

618 Citations (Scopus)

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 language	English
Journal	Nature Genetics
Volume	40
Issue number	11
Pages (from-to)	1375-83
Number of pages	8
ISSN	1061-4036
DOIs	https://doi.org/10.1038/ng.248
Publication status	Published - 2008

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title = "Single-copy insertion of transgenes in Caenorhabditis elegans.",

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.",

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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

SN - 1061-4036

VL - 40

SP - 1375

EP - 1383

JO - Nature Genetics

JF - Nature Genetics

IS - 11

ER -

Single-copy insertion of transgenes in Caenorhabditis elegans.

Abstract

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