Insertional Mutations in Transgenic Mice

Costantini, Frank; Radice, Glenn L.; Lee, James J.; Chada, Kiran; Perry, William L.; Son, Hyeung Jin

doi:10.1016/s0079-6603(08)60169-5

Cited by 56 publications

(23 citation statements)

References 22 publications

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“…These components need to be sequentially targeted into cells. Random integration of these elements into the genome is problematic because position effect variegation can profoundly affect expression and the insertion event itself may disrupt endogenous genes (Costantini et al, 1989;Palmiter and Brinster, 1986;Soriano et al, 1987).…”

mentioning

confidence: 99%

Efficient method to generate single‐copy transgenic mice by site‐specific integration in embryonic stem cells

Beard

Hochedlinger

Plath

et al. 2006

Genesis

456

580

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Summary: Transgenic and gene-targeted mutant mice provide powerful tools for analysis of the cellular processes involved in early development and in the pathogenesis of many diseases. Here we describe a transgene integration strategy mediated by site-specific recombination that allows establishment of multiple embryonic stem (ES) cell lines carrying tetracycline-inducible genes targeted to a specific locus to assure predictable temporal and spatial expression in ES cells and mice. Using homologous recombination we inserted an frt homing site into which tetracycline-inducible transgenes can be integrated efficiently in the presence of FLPe recombinase. This strategy and the vectors described here are generally applicable to any locus in ES cells and should allow for the rapid production of mice with transgenes efficiently targeted to a defined site.

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mentioning

confidence: 99%

Efficient method to generate single‐copy transgenic mice by site‐specific integration in embryonic stem cells

Beard

Hochedlinger

Plath

et al. 2006

Genesis

456

580

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“…DISCUSSION DNA-and retrovirus-mediated gene transfer has previously been used as a means of insertional mutagenesis in preimplantation embryos (34)(35)(36)(37)(38)(39)(40) and in ES cells (9,10,(14)(15)(16). As with vectors that rely on splicing to generate fusion transcripts, the U3LacZ promoter trap vector offers several advantages for disrupting genes in totipotent ES cells.…”

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confidence: 99%

Fluorescence-activated sorting of totipotent embryonic stem cells expressing developmentally regulated lacZ fusion genes.

Reddy

Rayburn

Melchner

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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Murine embryonic stem (ES) cells were infected with a retrovirus promoter trap vector, and clones expressing lacZ fusion genes (LacZ+) were isolated by fluorescence-activated cell sorting (FACS). Of 12 fusion genes tested, 1 was repressed when ES cells were allowed to differentiate in vitro. Two of three lacZ fusion genes tested were passed into the germ line, indicating that FACS does not significantly affect stem cell totipotency. The pattern of lacZ expression observed in vivo was consistent with that seen in vitro. Both fusion genes were expressed in preimplantation blastulas. However, a fusion gene whose expression was unaffected by in vitro differentiation was ubiquitously expressed in day-10 embryos, while the other, which showed regulated expression in vitro, was restricted to cells located along the posterior neural fold, the optic chiasm, and within the fourth ventricle. These results demonstrate the utility of using promoter trap vectors in conijunction with fluorescence sorting to disrupt developmentally regulated genes in mice.

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“…The integration of exogenous DNA sequences into the genome of mouse embryos can lead to insertional inactivation of endogenous genes (1)(2)(3). Insertional mutations may lead to a variety ofphenotypes such as embryonic lethalities (4)(5)(6)(7)(8) as well as morphological and neurological disorders (9)(10)(11)(12)(13).…”

mentioning

confidence: 99%

Symplastic spermatids (sys): a recessive insertional mutation in mice causing a defect in spermatogenesis.

MacGregor

Russell

Beek

et al. 1990

Proc. Natl. Acad. Sci. U.S.A.

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A line of transgenic mice that carries an insertional mutation in a gene essential for spermatogenesis is described. Males homozygous for the transgenic insert are sterile, while female homozygotes and both male and female heterozygotes exhibit normal fertility. Developing spermatids in homozygous males form prominent abnormal multinucleated syncytia (symplasts) and do not complete maturation. In addition, abnormal cytoplasmic vacuolation is commonly seen in Sertoli cells. One flank of the transgenic integration site within the genome has been cloned and used to show linkage between homozygosity for the transgene and the mutant phe-

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Insertional Mutations in Transgenic Mice

Cited by 56 publications

References 22 publications

Efficient method to generate single‐copy transgenic mice by site‐specific integration in embryonic stem cells

Efficient method to generate single‐copy transgenic mice by site‐specific integration in embryonic stem cells

Fluorescence-activated sorting of totipotent embryonic stem cells expressing developmentally regulated lacZ fusion genes.

Symplastic spermatids (sys): a recessive insertional mutation in mice causing a defect in spermatogenesis.

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