Conditional alleles in mice: Practical considerations for tissue‐specific knockouts

Kwan, Kin Ming

doi:10.1002/gene.10068

Cited by 149 publications

(110 citation statements)

References 135 publications

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“…In addition, the Cre/loxP and the Flp/FRT systems do not need any additional cofactors or sequence requirements for site-specific recombination in the mouse. Further reading about Cre-and Flp-based strategies for generating chromosomal rearrangements including large deletions and transloca-tions is provided by several excellent reviews (77,80,98,164).…”

Section: Conditional Gene Targetingmentioning

confidence: 99%

Of mice and models: improved animal models for biomedical research

Bockamp

Maringer

Spangenberg

et al. 2002

Physiological Genomics

147

109

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The ability to engineer the mouse genome has profoundly transformed biomedical research. During the last decade, conventional transgenic and gene knockout technologies have become invaluable experimental tools for modeling genetic disorders, assigning functions to genes, evaluating drugs and toxins, and by and large helping to answer fundamental questions in basic and applied research. In addition, the growing demand for more sophisticated murine models has also become increasingly evident. Good state-of-principle knowledge about the enormous potential of second-generation conditional mouse technology will be beneficial for any researcher interested in using these experimental tools. In this review we will focus on practice, pivotal principles, and progress in the rapidly expanding area of conditional mouse technology. The review will also present an internet compilation of available tetracycline-inducible mouse models as tools for biomedical research ( http://www.zmg.uni-mainz.de/tetmouse/ ).

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Section: Conditional Gene Targetingmentioning

confidence: 99%

Of mice and models: improved animal models for biomedical research

Bockamp

Maringer

Spangenberg

et al. 2002

Physiological Genomics

147

109

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“…In some instances conditional cell type selective knockouts had to be generated to circumvent embryonic lethality of global mouse knockouts [1][2][3]. Adipocyte selective inactivation of genes was possible through the identification of promoter sequences of the aP2 (fatty acid binding protein 4, FABP4) gene [4] and the introduction of Cre/loxP technology [5]. In recent years, these adipocyte restricted knockouts were also used with the particular purpose to define the function of a protein in these lipid storing cells, independent of its role in other tissues [6][7][8], or to uncover the function of the protein in mature adipocytes as opposed to its role during adipogenesis [9].…”

Section: Introductionmentioning

confidence: 99%

Ectopic recombination in the central and peripheral nervous system by aP2/FABP4‐Cre mice: Implications for metabolism research

2010

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Edited by Laszlo Nagy Keywords:Adipose tissue Conditional knockout mice Fatty acid binding protein 4 Sympathetic nervous system Peroxisomes Central nervous system Adipocyte a b s t r a c t aP2-Cre mice have amply been used to generate conditional adipose selective inactivation of important signaling molecules. We show that the efficiency of Cre mediated recombination in adipocytes and adipose selectivity is not always guaranteed. In particular, Cre activity was found in ganglia of the peripheral nervous system (PNS), in adrenal medulla and in neurons throughout the central nervous system (CNS). Because these tissues have an important impact on adipose tissue, care should be taken when using aP2-Cre mice to define the role of the targeted genes in adipose tissue function.

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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

316

389

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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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Conditional alleles in mice: Practical considerations for tissue‐specific knockouts

Cited by 149 publications

References 135 publications

Of mice and models: improved animal models for biomedical research

Of mice and models: improved animal models for biomedical research

Ectopic recombination in the central and peripheral nervous system by aP2/FABP4‐Cre mice: Implications for metabolism research

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

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