A systematic enhancer screen using lentivector transgenesis identifies conserved and non-conserved functional elements at the Olig1 and Olig2 locus

Article Article scientifique Article A systematic enhancer screen using lentivector transgenesis identifies conserved and non-conserved functional elements at the Olig1 and Olig2 locus eng 2010 author Friedli Marc 134865 author Barde Isabelle author Arcangeli Mélanie 538310 author Verp Sonia author Quazzola Alexandra 549267 author Zakany Jozsef 29602 author Lin-Marq Nathalie author Robyr Daniel 639042 author Attanasio Catia 103069 author Spitz François 508177 author Duboule Denis 28643 author Trono Didier 116006 author Antonarakis Stylianos 22813 e15741 10.1371/journal.pone.0015741 21206754 1932-6203 http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3012086/pdf/pone.0015741.pdf Finding sequences that control expression of genes is central to understanding genome function. Previous studies have used evolutionary conservation as an indicator of regulatory potential. Here, we present a method for the unbiased in vivo screen of putative enhancers in large DNA regions, using the mouse as a model. We cloned a library of 142 overlapping fragments from a 200 kb-long murine BAC in a lentiviral vector expressing LacZ from a minimal promoter, and used the resulting vectors to infect fertilized murine oocytes. LacZ staining of E11 embryos obtained by first using the vectors in pools and then testing individual candidates led to the identification of 3 enhancers, only one of which shows significant evolutionary conservation. In situ hybridization and 3C/4C experiments suggest that this enhancer, which is active in the neural tube and posterior diencephalon, influences the expression of the Olig1 and/or Olig2 genes. This work provides a new approach for the large-scale in vivo screening of transcriptional regulatory sequences, and further demonstrates that evolutionary conservation alone seems too limiting a criterion for the identification of enhancers. Animals Basic Helix-Loop-Helix Transcription Factors/*genetics Central Nervous System/metabolism Chickens Chromosomes, Artificial, Bacterial Enhancer Elements, Genetic Humans In Situ Hybridization Lac Operon Lentivirus/*genetics Mice Nerve Tissue Proteins/*genetics Promoter Regions, Genetic Sequence Analysis, DNA Transgenes Dewey 576.5 Dewey 590 44 Département de génétique et évolution 162 135 Département de médecine génétique et développement 21D false 196 Pathologie Moléculaire de la Trisomie 21 et le Génome Humain 248 PloS one 5 12 ca9b1b7b-4405-4101-9d05-e48e79ff960b Article (Published version) Public file1.pdf 1059825 application/pdf CC BY openAccess Faculté des sciences / Section de biologie / Département de génétique et évolution Faculté de médecine / Section de médecine fondamentale / Département de médecine génétique et développement

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A systematic enhancer screen using lentivector transgenesis identifies conserved and non-conserved functional elements at the Olig1 and Olig2 locus	PloS one			2010	865	490

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Quazzola, Alexandra