原创力文档a systematic enhancer screen using lentivector transgenesis identifies conserved and non-conserved functional elements at the olig1 and olig2 locus系统增强器屏幕使用lentivector转基因技术识别守恒和non-conserved通过简单地去除olig1和olig2轨迹的功能元素.pdfVIP
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a systematic enhancer screen using lentivector transgenesis identifies conserved and non-conserved functional elements at the olig1 and olig2 locus系统增强器屏幕使用lentivector转基因技术识别守恒和non-conserved通过简单地去除olig1和olig2轨迹的功能元素
A Systematic Enhancer Screen Using Lentivector
Transgenesis Identifies Conserved and Non-Conserved
Functional Elements at the Olig1 and Olig2 Locus
1,4 2,4 ´ 1,4 2,4 2,4
Marc Friedli , Isabelle Barde , Melanie Arcangeli , Sonia Verp , Alexandra Quazzola , Jozsef
Zakany3,4, Nathalie Lin-Marq1,4, Daniel Robyr1,4, Catia Attanasio1,4¤a, Franc¸ois Spitz3,4¤b, Denis
Duboule2,3,4*, Didier Trono2,4*, Stylianos E. Antonarakis1,4*
1 Department of Genetic Medicine and Development, University of Geneva Medical School and University Hospitals of Geneva, Geneva, Switzerland, 2 School of Life
´ ´
Sciences, Ecole Polytechnique Federale de Lausanne (EPFL), Lausanne, Switzerland, 3 Department of Zoology and Animal Biology, University of Geneva, Geneva,
Switzerland, 4 ‘‘Frontiers in Genetics’’, National Centre for Competence in Research, Bern, Switzerland
Abstract
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
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