Enhancer detection in the zebrafish using pseudotyped murine retroviruses.
Identifieur interne : 002142 ( PubMed/Curation ); précédent : 002141; suivant : 002143Enhancer detection in the zebrafish using pseudotyped murine retroviruses.
Auteurs : Mary Laplante [Norvège] ; Hiroshi Kikuta ; Melanie König ; Thomas S. BeckerSource :
- Methods (San Diego, Calif.) [ 1046-2023 ] ; 2006.
Descripteurs français
- KwdFr :
- Analyse de séquence d'ADN, Animal génétiquement modifié (génétique), Animaux, Biologie informatique, Clonage moléculaire, Danio zébré (embryologie), Danio zébré (génétique), Gènes rapporteurs, Génie génétique (), Génie génétique (instrumentation), Génomique (), Retroviridae (génétique), Souris, Transfection (), Transfection (instrumentation), Vecteurs génétiques, Éléments activateurs (génétique).
- MESH :
- embryologie : Danio zébré.
- génétique : Animal génétiquement modifié, Danio zébré, Retroviridae.
- instrumentation : Analyse de séquence d'ADN, Animaux, Biologie informatique, Clonage moléculaire, Gènes rapporteurs, Génie génétique, Génomique, Souris, Transfection, Vecteurs génétiques, Éléments activateurs (génétique).
English descriptors
- KwdEn :
- Animals, Animals, Genetically Modified (genetics), Cloning, Molecular, Computational Biology, Enhancer Elements, Genetic, Genes, Reporter, Genetic Engineering (instrumentation), Genetic Engineering (methods), Genetic Vectors, Genomics (methods), Mice, Retroviridae (genetics), Sequence Analysis, DNA, Transfection (instrumentation), Transfection (methods), Zebrafish (embryology), Zebrafish (genetics).
- MESH :
- embryology : Zebrafish.
- genetics : Animals, Genetically Modified, Retroviridae, Zebrafish.
- instrumentation : Genetic Engineering, Transfection.
- methods : Genetic Engineering, Genomics, Transfection.
- Animals, Cloning, Molecular, Computational Biology, Enhancer Elements, Genetic, Genes, Reporter, Genetic Vectors, Mice, Sequence Analysis, DNA.
Abstract
Vectors based on murine retroviruses are among the most efficient means to insert reporter constructs into the context of a vertebrate chromosome with the aim to visualize cis-regulatory information available to a basal promoter at the site of insertion. In combination with using the zebrafish embryo as a readout for the activity of regulatory elements, enhancer detection becomes a powerful technique for gene discovery and for the mapping of the extent of regulatory domains in a vertebrate genome. Our laboratory has performed the only large-scale enhancer detection screen to date in any vertebrate and we describe in this paper the methods we developed to generate viral particles, to insert reporter constructs into the zebrafish germ line, the screening of detection events in heterozygous F1 embryos, and the isolation of genomic sequence flanking the inserted vector for the purpose of genomic mapping. Given sufficient scale, the technology described here can be used to obtain cis-regulatory information across the entire zebrafish genome for any given basal promoter.
DOI: 10.1016/j.ymeth.2006.01.003
PubMed: 16887366
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pubmed:16887366Le document en format XML
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<term>Animals, Genetically Modified (genetics)</term>
<term>Cloning, Molecular</term>
<term>Computational Biology</term>
<term>Enhancer Elements, Genetic</term>
<term>Genes, Reporter</term>
<term>Genetic Engineering (instrumentation)</term>
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<term>Sequence Analysis, DNA</term>
<term>Transfection (instrumentation)</term>
<term>Transfection (methods)</term>
<term>Zebrafish (embryology)</term>
<term>Zebrafish (genetics)</term>
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<term>Animal génétiquement modifié (génétique)</term>
<term>Animaux</term>
<term>Biologie informatique</term>
<term>Clonage moléculaire</term>
<term>Danio zébré (embryologie)</term>
<term>Danio zébré (génétique)</term>
<term>Gènes rapporteurs</term>
<term>Génie génétique ()</term>
<term>Génie génétique (instrumentation)</term>
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<term>Retroviridae (génétique)</term>
<term>Souris</term>
<term>Transfection ()</term>
<term>Transfection (instrumentation)</term>
<term>Vecteurs génétiques</term>
<term>Éléments activateurs (génétique)</term>
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<keywords scheme="MESH" qualifier="embryologie" xml:lang="fr"><term>Danio zébré</term>
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<keywords scheme="MESH" qualifier="embryology" xml:lang="en"><term>Zebrafish</term>
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<keywords scheme="MESH" qualifier="genetics" xml:lang="en"><term>Animals, Genetically Modified</term>
<term>Retroviridae</term>
<term>Zebrafish</term>
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<term>Retroviridae</term>
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<keywords scheme="MESH" qualifier="instrumentation" xml:lang="en"><term>Genetic Engineering</term>
<term>Transfection</term>
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<term>Genetic Vectors</term>
<term>Mice</term>
<term>Sequence Analysis, DNA</term>
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<term>Animaux</term>
<term>Biologie informatique</term>
<term>Clonage moléculaire</term>
<term>Gènes rapporteurs</term>
<term>Génie génétique</term>
<term>Génomique</term>
<term>Souris</term>
<term>Transfection</term>
<term>Vecteurs génétiques</term>
<term>Éléments activateurs (génétique)</term>
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<front><div type="abstract" xml:lang="en">Vectors based on murine retroviruses are among the most efficient means to insert reporter constructs into the context of a vertebrate chromosome with the aim to visualize cis-regulatory information available to a basal promoter at the site of insertion. In combination with using the zebrafish embryo as a readout for the activity of regulatory elements, enhancer detection becomes a powerful technique for gene discovery and for the mapping of the extent of regulatory domains in a vertebrate genome. Our laboratory has performed the only large-scale enhancer detection screen to date in any vertebrate and we describe in this paper the methods we developed to generate viral particles, to insert reporter constructs into the zebrafish germ line, the screening of detection events in heterozygous F1 embryos, and the isolation of genomic sequence flanking the inserted vector for the purpose of genomic mapping. Given sufficient scale, the technology described here can be used to obtain cis-regulatory information across the entire zebrafish genome for any given basal promoter.</div>
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<Abstract><AbstractText>Vectors based on murine retroviruses are among the most efficient means to insert reporter constructs into the context of a vertebrate chromosome with the aim to visualize cis-regulatory information available to a basal promoter at the site of insertion. In combination with using the zebrafish embryo as a readout for the activity of regulatory elements, enhancer detection becomes a powerful technique for gene discovery and for the mapping of the extent of regulatory domains in a vertebrate genome. Our laboratory has performed the only large-scale enhancer detection screen to date in any vertebrate and we describe in this paper the methods we developed to generate viral particles, to insert reporter constructs into the zebrafish germ line, the screening of detection events in heterozygous F1 embryos, and the isolation of genomic sequence flanking the inserted vector for the purpose of genomic mapping. Given sufficient scale, the technology described here can be used to obtain cis-regulatory information across the entire zebrafish genome for any given basal promoter.</AbstractText>
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