Targeted expression of Cre recombinase to adipose tissue of transgenic mice directs adipose-specific excision of loxP-flanked gene segments

Barlow, Carrolee; Schroeder, Mona; Lekstrom-Himes, Julie; Kylefjord, Helen; Deng, Chu Xia; Wynshaw‐Boris, Anthony; Spiegelman, Bruce M.; Xanthopoulos, Kleanthis G.

doi:10.1093/nar/25.12.2543

Cited by 59 publications

(30 citation statements)

References 13 publications

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“…An alternative strategy would be to combine conditional Rb knockout mice 46,47 with transgenic mice expressing the Cre recombinase specifically in fat from the aP2 promoter/enhancer. 48 Thereby, supposedly viable mice lacking pRB specifically in fat could be generated. A potential problem with this strategy is the timing of disappearance of pRB from the differentiating adipocytes.…”

Section: Is Prb a Regulator Of White Versus Brown Adipocyte Differentmentioning

confidence: 99%

Novel Function of the Retinoblastoma Protein in Fat: Regulation of White Versus Brown Adipocyte Differentiation

Hansen¹,

Riele²,

Kristiansen³

2004

Cell Cycle

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The differentiation of white and brown fat cells is controlled by a similar set of transcription factors, including PPARγ and C/EBPα. However, despite many similarities between the two types of fat cells, they carry out essentially opposite functions in vivo, with white adipocytes being the major energy store and brown adipocytes being potent energy-dissipaters through thermogenesis. Yet, little is known about factors differentially regulating the formation of white and brown fat cells. Members of the retinoblastoma protein family (pRB, p107, p130) have been implicated in the regulation of adipocyte differentiation, and expression and phosphorylation of the three retinoblastoma family proteins oscillate in a characteristic manner during differentiation of the white preadipocyte cell line 3T3-L1. We have recently demonstrated a surprising function of the retinoblastoma protein in the regulation of white versus brown adipocyte differentiation in vitro and possibly in vivo. Here we summarize the current knowledge on the retinoblastoma protein in fat cells, with particular emphasis on its potential role in adipocyte lineage commitment and differentiation. © 2 0 0 4 L a n d e s B i o s c i e n c e . D o N o t D i s t r i b u t e www.landesbioscience.com Cell Cycle REGULATION OF WHITE VERSUS BROWN ADIPOCYTE DIFFERENTIATION 776 Cell Cycle 2004; Vol. 3 Issue 6 © 2 0 0 4 L a n d e s B i o s c i e n c e . D o N o t D i s t r i b u t e www.landesbioscience.com Cell Cycle REGULATION OF WHITE VERSUS BROWN ADIPOCYTE DIFFERENTIATION

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Section: Is Prb a Regulator Of White Versus Brown Adipocyte Differentmentioning

confidence: 99%

Novel Function of the Retinoblastoma Protein in Fat: Regulation of White Versus Brown Adipocyte Differentiation

Hansen¹,

Riele²,

Kristiansen³

2004

Cell Cycle

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“…The Cre/loxP recombination system of bacteriophage P1 is commonly used for manipulation of transgenes, where the Cre recombinase directs both excision and insertion of DNA segments flanked by loxP sites in any cellular environment (Barlow et al 1997;Nagy 2000). Cre recombinase expression achieved by classic transgenesis or targeting to an appropriate locus may be tissuespecific, temporally restricted, or inducible (Soriano 1999).…”

Section: Introductionmentioning

confidence: 99%

Selective expression of an aP2/Fatty Acid Binding Protein4-Cre transgene in non-adipogenic tissues during embryonic development

et al. 2006

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Mouse strains expressing the site-specific Cre recombinase facilitate conditional ablation or activation of genomic sequences when one or several exons of a gene of interest are flanked by loxP sites. Recently, several strains targeting Cre expression to adipocytes have been developed using promoter sequences from the aP2 (Fatty Acid Binding Protein 4, FABP4) gene for adipose tissue-specific gene expression studies. aP2/FABP4 is predominantly expressed in adipose tissue, and while this promoter provides adipocyte-restricted expression postnatally, its expression throughout embryonic development had not been previously characterized. In this report, we demonstrate that the aP2-Cre transgene is expressed and consistently localized within the embryo from mid-gestation stage 9.5 dpc. By 15.5 dpc, beta-gal activity was detected primarily in the brown adipose tissue, trigeminal ganglia, dorsal root ganglia, cartilage primordia and vertebrae. Immunofluorescence staining for Cre recombinase and FABP4 protein showed a corresponding staining pattern similar to that of beta-gal, confirming that Cre recombinase was produced in the transgenic line at late stages of development, and overlapped with endogenous aP2/FABP4 production. Further, fat-specific oil red O staining of tissue sections validated the presence of lipids in the stained tissues indicating that adipocytes and/or adipocyte-like cells were indeed present in these tissues. This is the first report to our knowledge to describe and confirm aP2/FABP4 promoter expression in this transgenic line during development in the mouse embryo and indicates that aP2/FABP4 expression occurs not only in mature adipocytes, but has a wider embryonic expression pattern than previously appreciated.

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“…Indeed, this Cre recombinase efficiently can excise a DNA segment flanked by two loxP sites (floxed DNA) in animal cells (5). Placing the Cre gene under the control of either a cell-specific (6)(7)(8) or an inducible (9) promoter can lead, through the excision of the floxed DNA segment, to either spatially or temporally controlled somatic mutations, respectively. However, these conditional gene targeting systems for generating somatic mutations have also a number of limitations because they are only spatially or temporally controlled.…”

mentioning

confidence: 99%

Spatio-temporally controlled site-specific somatic mutagenesis in the mouse

Brocard

Warot

Wendling

et al. 1997

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

222

142

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The efficient introduction of somatic mutations in a given gene, at a given time, in a specific cell type will facilitate studies of gene function and the generation of animal models for human diseases. We have shown previously that conditional recombination-excision between two loxP sites can be achieved in mice by using the Cre recombinase fused to a mutated ligand binding domain of the human estrogen receptor (Cre-ER T ), which binds tamoxifen but not estrogens. DNA excision was induced in a number of tissues after administration of tamoxifen to transgenic mice expressing Cre-ER T under the control of the cytomegalovirus promoter. However, the efficiency of excision varied between tissues, and the highest level (Ϸ40%) was obtained in the skin. To determine the efficiency of excision mediated by Cre-ER T in a given cell type, we have now crossed Cre-ER T -expressing mice with reporter mice in which expression of Escherichia coli ␤-galactosidase can be induced through Cre-mediated recombination. The efficiency and kinetics of this recombination were analyzed at the cellular level in the epidermis of 6-to 8-week-old double transgenic mice. We show that site-specific excision occurred within a few days of tamoxifen treatment in essentially all epidermis cells expressing Cre-ER T . These results indicate that cell-specific expression of Cre-ER T in transgenic mice can be used for efficient tamoxifendependent, Cre-mediated recombination at loci containing loxP sites to generate site-specific somatic mutations in a spatio-temporally controlled manner.

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Targeted expression of Cre recombinase to adipose tissue of transgenic mice directs adipose-specific excision of loxP-flanked gene segments

Cited by 59 publications

References 13 publications

Novel Function of the Retinoblastoma Protein in Fat: Regulation of White Versus Brown Adipocyte Differentiation

Novel Function of the Retinoblastoma Protein in Fat: Regulation of White Versus Brown Adipocyte Differentiation

Selective expression of an aP2/Fatty Acid Binding Protein4-Cre transgene in non-adipogenic tissues during embryonic development

Spatio-temporally controlled site-specific somatic mutagenesis in the mouse

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