Kinetics and epigenetics of retroviral silencing in mouse embryonic stem cells defined by deletion of the D4Z4 element
| Autor: | Mandy Y. M. Lo, James Ellis, Matthew C. Lorincz, Peter Pasceri, Sylvie Rival-Gervier, Shahryar Khattak |
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| Přispěvatelé: | Biologie du développement et reproduction (BDR), Centre National de la Recherche Scientifique (CNRS)-École nationale vétérinaire d'Alfort (ENVA)-Institut National de la Recherche Agronomique (INRA), Developmental and Stem Cell Biology, Hospital for Sick Children, Department of Molecular Genetics, University of Toronto, Department of Medical Genetics, Life Sciences Institute, University of British Columbia (UBC), Canadian Institutes of Health Research, CIHR MOP-111065, IG1-102956 RMF-92090, Biologie du Développement et Reproduction (BDR), Institut National de la Recherche Agronomique (INRA) |
| Rok vydání: | 2012 |
| Předmět: |
souris
Epigenesis Genetic Histones Mice 0302 clinical medicine Proviruses Drug Discovery Gene Order Transgenes Homologous Recombination [SDV.BDD]Life Sciences [q-bio]/Development Biology Epigenomics Sequence Deletion Regulation of gene expression Genetics 0303 health sciences cinétique culture cellulaire expression de transgène Biologie du développement Development Biology Long terminal repeat Chromatin DNA methylation Molecular Medicine Original Article Insulator Elements cellule souche embryonnaire expression génique Heterochromatin Genetic Vectors Biology Methylation 03 medical and health sciences épigénétique Gene silencing Animals Epigenetics Gene Silencing Molecular Biology Embryonic Stem Cells 030304 developmental biology Pharmacology Terminal Repeat Sequences DNA Methylation Kinetics Retroviridae Gene Expression Regulation Commentary vecteur rétroviral 030217 neurology & neurosurgery |
| Zdroj: | Molecular Therapy 8 (21), 1536-1550. (2013) Molecular Therapy Molecular Therapy, Cell Press, 2013, 21 (8), pp.1536-1550. ⟨10.1038/mt.2013.131⟩ Molecular Therapy, Nature Publishing Group, 2013, 21 (8), pp.1536-1550. ⟨10.1038/mt.2013.131⟩ |
| ISSN: | 1525-0024 1525-0016 |
| DOI: | 10.1038/mt.2013.131⟩ |
| Popis: | Retroviral vectors are silenced in embryonic stem (ES) cells by epigenetic mechanisms whose kinetics are poorly understood. We show here that a 3'D4Z4 insulator directs retroviral expression with persistent but variable expression for up to 5 months. Combining an internal 3'D4Z4 with HS4 insulators in the long terminal repeats (LTRs) shows that these elements cooperate, and defines the first retroviral vector that fully escapes long-term silencing. Using FLP recombinase to induce deletion of 3'D4Z4 from the provirus in ES cell clones, we established retroviral silencing at many but not all integration sites. This finding shows that 3'D4Z4 does not target retrovirus integration into favorable epigenomic domains but rather protects the transgene from silencing. Chromatin analyses demonstrate that 3'D4Z4 blocks the spread of heterochromatin marks including DNA methylation and repressive histone modifications such as H3K9 methylation. In addition, our deletion system reveals three distinct kinetic classes of silencing (rapid, gradual or not silenced), in which multiple epigenetic pathways participate in silencing at different integration sites. We conclude that vectors with both 3'D4Z4 and HS4 insulator elements fully block silencing, and may have unprecedented utility for gene transfer applications that require long-term gene expression in pluripotent stem (PS) cells. |
| Databáze: | OpenAIRE |
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