Conditional mutagenesis in mice with heat shock promoter-driven cre transgenes

Paula Dietrich, Ioannis Dragatsis, Shouhong Xuan, Scott Zeitlin, Argiris Efstratiadis

Research output: Contribution to journalArticle

85 Citations (Scopus)

Abstract

To explore the potential of a simple and rapid approach for ubiquitous conditional gene disruption, we have generated Cre-producer mouse transgenic lines (Hs-cre1, 6 and 7) expressing a recombinase transgene (cre) from a heat shock gene promoter and tested their performance in Cre-mediated excision of target DNA in crosses with Cre-responder strains carrying loxP- modified alleles of the genes encoding the Huntington's disease gene homolog (Hdh), the epidermal growth factor receptor (Egfr), and the type 1 insulin- like growth factor receptor (Igflr). Analyses of progeny possessing various transgene/reporter combinations showed that cre expression can occur without heat shock in early embryos, but this constitutive transcription is stochastic and transgene dependent. Thus, Hs-cre1 behaves predominantly as a 'deleter' strain, since the majority of progeny (~70-85%) exhibit complete recombination, regardless of reporter locus. Lines Hs-cre6 and Hs-cre7, however, function successfully as 'mosaicking' strains because, in addition to two extreme classes of progeny with 0% or 100% recombination, they generate an intermediate class of mosaics exhibiting various degrees of partial DNA excision. Notably, the frequency of offspring in each class varies between reporters, but mosaic embryos are consistently obtained in adequate numbers (~30-60%). The Hs-cre6 transgene is also inducible and can be used to introduce mosaicism into adult tissues at preselected developmental times by heat shock treatment of mice with 0% recombination in tail DNA. By bypassing the lethality resulting from some gene knockouts, mosaic embryos and mice make particular mutational analyses possible and are also very useful for the identification of cell lineage-specific gene functions.

Original languageEnglish (US)
Pages (from-to)196-205
Number of pages10
JournalMammalian Genome
Volume11
Issue number3
DOIs
StatePublished - Mar 15 2000

Fingerprint

Transgenes
Mutagenesis
Shock
Hot Temperature
Genetic Recombination
Embryonic Structures
Genes
DNA
erbB-1 Genes
IGF Type 1 Receptor
Gene Knockout Techniques
Mosaicism
Huntington Disease
Cell Lineage
Transgenic Mice
Tail
Alleles

All Science Journal Classification (ASJC) codes

  • Genetics

Cite this

Conditional mutagenesis in mice with heat shock promoter-driven cre transgenes. / Dietrich, Paula; Dragatsis, Ioannis; Xuan, Shouhong; Zeitlin, Scott; Efstratiadis, Argiris.

In: Mammalian Genome, Vol. 11, No. 3, 15.03.2000, p. 196-205.

Research output: Contribution to journalArticle

Dietrich, Paula ; Dragatsis, Ioannis ; Xuan, Shouhong ; Zeitlin, Scott ; Efstratiadis, Argiris. / Conditional mutagenesis in mice with heat shock promoter-driven cre transgenes. In: Mammalian Genome. 2000 ; Vol. 11, No. 3. pp. 196-205.
@article{8b396e05d6694909b2cd481376f1b449,
title = "Conditional mutagenesis in mice with heat shock promoter-driven cre transgenes",
abstract = "To explore the potential of a simple and rapid approach for ubiquitous conditional gene disruption, we have generated Cre-producer mouse transgenic lines (Hs-cre1, 6 and 7) expressing a recombinase transgene (cre) from a heat shock gene promoter and tested their performance in Cre-mediated excision of target DNA in crosses with Cre-responder strains carrying loxP- modified alleles of the genes encoding the Huntington's disease gene homolog (Hdh), the epidermal growth factor receptor (Egfr), and the type 1 insulin- like growth factor receptor (Igflr). Analyses of progeny possessing various transgene/reporter combinations showed that cre expression can occur without heat shock in early embryos, but this constitutive transcription is stochastic and transgene dependent. Thus, Hs-cre1 behaves predominantly as a 'deleter' strain, since the majority of progeny (~70-85{\%}) exhibit complete recombination, regardless of reporter locus. Lines Hs-cre6 and Hs-cre7, however, function successfully as 'mosaicking' strains because, in addition to two extreme classes of progeny with 0{\%} or 100{\%} recombination, they generate an intermediate class of mosaics exhibiting various degrees of partial DNA excision. Notably, the frequency of offspring in each class varies between reporters, but mosaic embryos are consistently obtained in adequate numbers (~30-60{\%}). The Hs-cre6 transgene is also inducible and can be used to introduce mosaicism into adult tissues at preselected developmental times by heat shock treatment of mice with 0{\%} recombination in tail DNA. By bypassing the lethality resulting from some gene knockouts, mosaic embryos and mice make particular mutational analyses possible and are also very useful for the identification of cell lineage-specific gene functions.",
author = "Paula Dietrich and Ioannis Dragatsis and Shouhong Xuan and Scott Zeitlin and Argiris Efstratiadis",
year = "2000",
month = "3",
day = "15",
doi = "10.1007/s003350010037",
language = "English (US)",
volume = "11",
pages = "196--205",
journal = "Mammalian Genome",
issn = "0938-8990",
publisher = "Springer New York",
number = "3",

}

TY - JOUR

T1 - Conditional mutagenesis in mice with heat shock promoter-driven cre transgenes

AU - Dietrich, Paula

AU - Dragatsis, Ioannis

AU - Xuan, Shouhong

AU - Zeitlin, Scott

AU - Efstratiadis, Argiris

PY - 2000/3/15

Y1 - 2000/3/15

N2 - To explore the potential of a simple and rapid approach for ubiquitous conditional gene disruption, we have generated Cre-producer mouse transgenic lines (Hs-cre1, 6 and 7) expressing a recombinase transgene (cre) from a heat shock gene promoter and tested their performance in Cre-mediated excision of target DNA in crosses with Cre-responder strains carrying loxP- modified alleles of the genes encoding the Huntington's disease gene homolog (Hdh), the epidermal growth factor receptor (Egfr), and the type 1 insulin- like growth factor receptor (Igflr). Analyses of progeny possessing various transgene/reporter combinations showed that cre expression can occur without heat shock in early embryos, but this constitutive transcription is stochastic and transgene dependent. Thus, Hs-cre1 behaves predominantly as a 'deleter' strain, since the majority of progeny (~70-85%) exhibit complete recombination, regardless of reporter locus. Lines Hs-cre6 and Hs-cre7, however, function successfully as 'mosaicking' strains because, in addition to two extreme classes of progeny with 0% or 100% recombination, they generate an intermediate class of mosaics exhibiting various degrees of partial DNA excision. Notably, the frequency of offspring in each class varies between reporters, but mosaic embryos are consistently obtained in adequate numbers (~30-60%). The Hs-cre6 transgene is also inducible and can be used to introduce mosaicism into adult tissues at preselected developmental times by heat shock treatment of mice with 0% recombination in tail DNA. By bypassing the lethality resulting from some gene knockouts, mosaic embryos and mice make particular mutational analyses possible and are also very useful for the identification of cell lineage-specific gene functions.

AB - To explore the potential of a simple and rapid approach for ubiquitous conditional gene disruption, we have generated Cre-producer mouse transgenic lines (Hs-cre1, 6 and 7) expressing a recombinase transgene (cre) from a heat shock gene promoter and tested their performance in Cre-mediated excision of target DNA in crosses with Cre-responder strains carrying loxP- modified alleles of the genes encoding the Huntington's disease gene homolog (Hdh), the epidermal growth factor receptor (Egfr), and the type 1 insulin- like growth factor receptor (Igflr). Analyses of progeny possessing various transgene/reporter combinations showed that cre expression can occur without heat shock in early embryos, but this constitutive transcription is stochastic and transgene dependent. Thus, Hs-cre1 behaves predominantly as a 'deleter' strain, since the majority of progeny (~70-85%) exhibit complete recombination, regardless of reporter locus. Lines Hs-cre6 and Hs-cre7, however, function successfully as 'mosaicking' strains because, in addition to two extreme classes of progeny with 0% or 100% recombination, they generate an intermediate class of mosaics exhibiting various degrees of partial DNA excision. Notably, the frequency of offspring in each class varies between reporters, but mosaic embryos are consistently obtained in adequate numbers (~30-60%). The Hs-cre6 transgene is also inducible and can be used to introduce mosaicism into adult tissues at preselected developmental times by heat shock treatment of mice with 0% recombination in tail DNA. By bypassing the lethality resulting from some gene knockouts, mosaic embryos and mice make particular mutational analyses possible and are also very useful for the identification of cell lineage-specific gene functions.

UR - http://www.scopus.com/inward/record.url?scp=0034007328&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034007328&partnerID=8YFLogxK

U2 - 10.1007/s003350010037

DO - 10.1007/s003350010037

M3 - Article

VL - 11

SP - 196

EP - 205

JO - Mammalian Genome

JF - Mammalian Genome

SN - 0938-8990

IS - 3

ER -