Conditional control of gene expression in the mouse

Lewandoski, Mark

doi:10.1038/35093537

Cited by 715 publications

(487 citation statements)

References 155 publications

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“…SLICK is based on the popular cre/loxP system that has been used extensively to achieve conditional gene knockout in mice 17,34,35 . The availability of mice harboring conditional knockout alleles is continually increasing and a large scale project to generate "floxed" conditional knockout alleles is underway 1 .…”

Section: Discussionmentioning

confidence: 99%

Single-neuron labeling with inducible Cre-mediated knockout in transgenic mice

et al. 2008

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To facilitate functional analysis of neuronal connectivity in a mammalian nervous system tightly packed with billions of cells, we developed a new technique that allows inducible genetic manipulations within fluorescently labeled single neurons in mice. We term this technique SLICK for Single-neuron Labeling with Inducible Cre-mediated Knockout. SLICK is achieved by coexpressing a drug-inducible form of cre recombinase and a fluorescent protein within the same small subsets of neurons. Thus, SLICK combines the powerful cre recombinase system for conditional genetic manipulation and the fluorescent labeling of single neurons for imaging. We demonstrate efficient inducible genetic manipulation in several types of neurons using SLICK. Furthermore, we apply SLICK to eliminate synaptic transmission in a small subset of neuromuscular junctions. Our results provide evidence for the long-term stability of inactive neuromuscular synapses in adult animals. More broadly, these studies demonstrate a cre-LoxP compatible system for dissecting gene functions in single identifiable neurons.

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Section: Discussionmentioning

confidence: 99%

Single-neuron labeling with inducible Cre-mediated knockout in transgenic mice

et al. 2008

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“…More recently, conditional gene targeting using the Cre/loxP system has emerged as an especially powerful method in reproductive genetics, particularly in the study of genes whose systemic inactivation is lethal. Critical to the success this approach is the availability of suitable Cre transgenes expressed in a wide range of distinct cell lineages at appropriate developmental timepoints (Kwan, 2002;Lewandoski, 2001). Previous germ cell Cre lines include ZP3-Cre, which is not active in primordial follicles but becomes induced following follicle growth; this line is thus useful for analyses of follicle growth or maternal contributions to the embryo (de Vries et al, 2000).…”

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confidence: 99%

Generation of a germ cell‐specific mouse transgenic Cre line, Vasa‐Cre

Gallardo

Shirley

John

et al. 2007

Genesis

315

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SummaryCell type-specific genetic modification using the Cre/loxP system is a powerful tool for genetic analysis of distinct cell lineages. Because of the exquisite specificity of Vasa expression (confined to the germ cell lineage in invertebrate and vertebrate species), we hypothesized that a Vasa promoterdriven transgenic Cre line would prove useful for the germ cell lineage-specific inactivation of genes. Here we describe a transgenic mouse line, Vasa-Cre, where Cre is efficiently and specifically expressed in germ cells. Northern analysis showed that transgene expression was confined to the gonads. Cre-mediated recombination with the Rosa26-lacZ reporter was observed beginning at ~e15, and was >95% efficient in male and female germ cells by birth. There was no ectopic activity in most adults, although some animals showed more widespread lacZ expression. This Vasa-Cre transgenic line should thus prove useful for genetic analysis of diverse aspects of germ cell function and gametogenesis. KeywordsCre; vasa; germ cells; gonad Genetic approaches are well-suited for studies of gametogenesis and reproduction, processes that require complex interactions between germline and somatic cells, and hence are difficult to fully recapitulate in vitro. More recently, conditional gene targeting using the Cre/loxP system has emerged as an especially powerful method in reproductive genetics, particularly in the study of genes whose systemic inactivation is lethal. Critical to the success this approach is the availability of suitable Cre transgenes expressed in a wide range of distinct cell lineages at appropriate developmental timepoints (Kwan, 2002;Lewandoski, 2001). Previous germ cell Cre lines include ZP3-Cre, which is not active in primordial follicles but becomes induced following follicle growth; this line is thus useful for analyses of follicle growth or maternal contributions to the embryo (de Vries et al., 2000). A Cre knock-in into the TNAP (tissue nonspecific alkaline phosphatase) is active in primordial germ cells starting as early as e9.5. but the TNAP promoter is also active in other tissues (e.g. placenta and liver, among others) and such widespread expression may complicate interpretation of experiments where germ cellspecific gene inactivation is desired or required (if e.g. the locus is essential) (Lomeli et al., 2000).Vasa expression is confined to the germ cell lineage in species including Drosophila, zebrafish, mouse, and humans (Castrillon et al., 2000;Lasko and Ashburner, 1988 (Tanaka et al., 2000). We reasoned that a Vasa-Cre line could prove useful for studies of germ cell function following gonadal colonization, including many aspects of spermatogenesis and oogenesis, such as the assembly, activation, and growth of primordial follicles.We cloned a 5.6 kb DNA fragment containing mouse Vasa 5' regulatory sequences, including the transcriptional start site. This fragment contains canonical promoter sequences including a TATA box at −27 and more distant control elements important for germ cell specific ex...

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“…These data demonstrate an important role for C/EBPb in keratinocytes survival and skin tumorigenesis. However, phenotypes of germline knockouts can be complicated by pleiotropic effects of the protein under study in various tissues and organs (Lewandoski, 2001). For example, germline deletion of Rb results in severe defects in neurogenesis involving increased apoptosis.…”

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confidence: 99%

Conditional ablation of C/EBPβ demonstrates its keratinocyte-specific requirement for cell survival and mouse skin tumorigenesis

et al. 2005

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The CCAAT/enhancer binding protein b (C/EBPb) is implicated in the regulation of many different molecular and physiological processes. Mice with a germline deletion of C/EBPb (C/EBPb À/À ) display phenotypes in a multitude of cell types and organ systems, including skin where C/EBPb À/À mice exhibit increased apoptosis in epidermal keratinocytes in response to carcinogen treatment and are completely resistant to carcinogen-induced skin tumorigenesis. To determine the contribution of systemic versus cell autonomous functions of C/EBPb to specific phenotypes, mice with a conditional 'floxed' C/EBPb null allele were generated. Epidermal-specific deletion of C/EBPb was achieved by Cre recombinase expression from a keratin 5 (K5) promoter. Similar to C/EBPb À/À mice, K5-Cre;C/ EBPb fl/fl mice were completely refractory to 7,12 dimethylbenz[a]anthracene (DMBA)-induced skin tumorigenesis and these mice displayed increased DMBA-induced apoptosis in epidermal keratinocytes compared to wild-type mice. In contrast, mice lacking the related gene, C/EBPd, were not resistant to DMBA-induced skin tumorigenesis, indicating a unique role of C/EBPb in skin tumor development. Our findings demonstrate that C/EBPb exerts an essential, keratinocyte-intrinsic role in cell survival in response to carcinogen treatment and the elimination of C/EBPb in keratinocytes is sufficient to confer complete resistance of the skin to chemical carcinogenesis.

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Conditional control of gene expression in the mouse

Cited by 715 publications

References 155 publications

Single-neuron labeling with inducible Cre-mediated knockout in transgenic mice

Single-neuron labeling with inducible Cre-mediated knockout in transgenic mice

Generation of a germ cell‐specific mouse transgenic Cre line, Vasa‐Cre

Conditional ablation of C/EBPβ demonstrates its keratinocyte-specific requirement for cell survival and mouse skin tumorigenesis

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