title: Construction and validation of a novel dual reporter vector for studying mammalian bidirectional promoters.
abstract: Regulation of gene expression plays important role in cellular functions. With the development of sequencing techniques, more and more genomes are available and genome-wide analyses of genomic structures that may affect gene expression regulation are now possible. Analyses of several genomes have found a class of regulatory regions that contain elements that initiate transcription of two different genes positioned with a head-to-head arrangement in two opposite directions. These regulatory regions are known as bidirectional promoters. Although bidirectional promoters have been known for years, recent genome-scale studies have shown that the regulation of the expression of up to 10% of the genes are controlled by bidirectional promoters. These findings are based mostly on computational work and only a limited number of putative bidirectional promoters have been experimentally validated. Developing methods to study bidirectional promoters will allow researchers to understand how these regions are regulated and the roles that divergent transcription plays in the expression of genes. Here, we have developed a novel dual-fluorescence reporter gene vector to study the transcriptional output of mammalian bidirectional promoters. We demonstrate that this vector is capable of expressing reporter genes under the control of bidirectional promoters, using the known human OSGEP/APEX bidirectional promoter.
authors.affiliation: INRA, UMR1313, Unité Génétique Animale et Biologie Intégrative, Domaine de Vilvert, F-78352 Jouy-en-Josas, France; AgroParisTech, UMR1313, Unité Génétique Animale et Biologie Intégrative, Domaine de Vilvert, F-78352 Jouy-en-Josas, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Léjard
authors.affiliation: INRA, UMR1313, Unité Génétique Animale et Biologie Intégrative, Domaine de Vilvert, F-78352 Jouy-en-Josas, France; AgroParisTech, UMR1313, Unité Génétique Animale et Biologie Intégrative, Domaine de Vilvert, F-78352 Jouy-en-Josas, France.
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Rebours
authors.affiliation: INRA, UMR1313, Unité Génétique Animale et Biologie Intégrative, Domaine de Vilvert, F-78352 Jouy-en-Josas, France; AgroParisTech, UMR1313, Unité Génétique Animale et Biologie Intégrative, Domaine de Vilvert, F-78352 Jouy-en-Josas, France.
authors.forename: Cédric
authors.initials: C
authors.lastname: Meersseman
authors.affiliation: INRA, UMR1313, Unité Génétique Animale et Biologie Intégrative, Domaine de Vilvert, F-78352 Jouy-en-Josas, France; AgroParisTech, UMR1313, Unité Génétique Animale et Biologie Intégrative, Domaine de Vilvert, F-78352 Jouy-en-Josas, France. Electronic address: dominique.rocha@jouy.inra.fr.
authors.forename: Dominique
authors.initials: D
authors.lastname: Rocha
mesh_terms: D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D019295:Computational Biology; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D056726:Genetic Association Studies / Q000379:methods; D005822:Genetic Vectors; D016678:Genome; D006801:Humans; D008322:Mammals / Q000235:genetics*; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D015203:Reproducibility of Results; D014158:Transcription, Genetic
chemical_list: D008666:Metalloendopeptidases; C070738:O-sialoglycoprotein endopeptidase
keywords: Bidirectional promoter; Flow cytometry; Fluorescent proteins; Reporter gene