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Mol Ther
2009 Aug 01;178:1434-41. doi: 10.1038/mt.2009.74.
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The sea urchin sns5 insulator protects retroviral vectors from chromosomal position effects by maintaining active chromatin structure.
D'Apolito D
,
Baiamonte E
,
Bagliesi M
,
Di Marzo R
,
Calzolari R
,
Ferro L
,
Franco V
,
Spinelli G
,
Maggio A
,
Acuto S
.
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Silencing and position-effect (PE) variegation (PEV), which is due to integration of viral vectors in heterochromatin regions, are considered significant obstacles to obtaining a consistent level of transgene expression in gene therapy. The inclusion of chromatin insulators into vectors has been proposed to counteract this position-dependent variegation of transgene expression. Here, we show that the sea urchin chromatin insulator, sns5, protects a recombinant gamma-retroviral vector from the negative influence of chromatin in erythroid milieu. This element increases the probability of vector expression at different chromosomal integration sites, which reduces both silencing and PEV. By chromatin immunoprecipitation (ChIP) analysis, we demonstrated the specific binding of GATA1 and OCT1 transcription factors and the enrichment of hyperacetylated nucleosomes to sns5 sequences. The results suggest that this new insulator is able to maintain a euchromatin state inside the provirus locus with mechanisms that are common to other characterized insulators. On the basis of its ability to function as barrier element in erythroid milieu and to bind the erythroid specific factor GATA1, the inclusion of sns5 insulator in viral vectors may be of practical benefit in gene transfer applications and, in particular, for gene therapy of erythroid disorders.
Acuto,
Functional characterization of the sea urchin sns chromatin insulator in erythroid cells.
2005, Pubmed,
Echinobase
Acuto,
Functional characterization of the sea urchin sns chromatin insulator in erythroid cells.
2005,
Pubmed
,
Echinobase
Aker,
Integration bias of gammaretrovirus vectors following transduction and growth of primary mouse hematopoietic progenitor cells with and without selection.
2006,
Pubmed
Arumugam,
Improved human beta-globin expression from self-inactivating lentiviral vectors carrying the chicken hypersensitive site-4 (cHS4) insulator element.
2007,
Pubmed
Bell,
Stopped at the border: boundaries and insulators.
1999,
Pubmed
Bell,
Insulators and boundaries: versatile regulatory elements in the eukaryotic genome.
2001,
Pubmed
Bernstein,
Genomic maps and comparative analysis of histone modifications in human and mouse.
2005,
Pubmed
Blobel,
CREB-binding protein and p300: molecular integrators of hematopoietic transcription.
2000,
Pubmed
Cheng,
A GFP reporter system to assess gene transfer and expression in human hematopoietic progenitor cells.
1997,
Pubmed
Di Caro,
Down-regulation of early sea urchin histone H2A gene relies on cis regulative sequences located in the 5' and 3' regions and including the enhancer blocker sns.
2004,
Pubmed
,
Echinobase
Di Caro,
Constitutive promoter occupancy by the MBF-1 activator and chromatin modification of the developmental regulated sea urchin alpha-H2A histone gene.
2007,
Pubmed
,
Echinobase
Di Simone,
The sea urchin sns insulator blocks CMV enhancer following integration in human cells.
2001,
Pubmed
,
Echinobase
Emery,
A chromatin insulator protects retrovirus vectors from chromosomal position effects.
2000,
Pubmed
Emery,
Development of virus vectors for gene therapy of beta chain hemoglobinopathies: flanking with a chromatin insulator reduces gamma-globin gene silencing in vivo.
2002,
Pubmed
Enver,
Developmental regulation of fetal to adult globin gene switching in human fetal erythroid x mouse erythroleukemia cell hybrids.
1991,
Pubmed
Gaszner,
Insulators: exploiting transcriptional and epigenetic mechanisms.
2006,
Pubmed
Geyer,
Protecting against promiscuity: the regulatory role of insulators.
2002,
Pubmed
Hacein-Bey-Abina,
LMO2-associated clonal T cell proliferation in two patients after gene therapy for SCID-X1.
2003,
Pubmed
Hantzopoulos,
Improved gene expression upon transfer of the adenosine deaminase minigene outside the transcriptional unit of a retroviral vector.
1989,
Pubmed
Hawley,
Versatile retroviral vectors for potential use in gene therapy.
1994,
Pubmed
Huang,
USF1 recruits histone modification complexes and is critical for maintenance of a chromatin barrier.
2007,
Pubmed
Karpen,
Position-effect variegation and the new biology of heterochromatin.
1994,
Pubmed
Kim,
A high-resolution map of active promoters in the human genome.
2005,
Pubmed
Kuhn,
Genomic insulators: connecting properties to mechanism.
2003,
Pubmed
Letting,
Formation of a tissue-specific histone acetylation pattern by the hematopoietic transcription factor GATA-1.
2003,
Pubmed
Li,
The cHS4 chromatin insulator reduces gammaretroviral vector silencing by epigenetic modifications of integrated provirus.
2008,
Pubmed
Litt,
Correlation between histone lysine methylation and developmental changes at the chicken beta-globin locus.
2001,
Pubmed
Margueron,
The key to development: interpreting the histone code?
2005,
Pubmed
Markowitz,
Construction and use of a safe and efficient amphotropic packaging cell line.
1988,
Pubmed
Melfi,
Functional characterization of the enhancer blocking element of the sea urchin early histone gene cluster reveals insulator properties and three essential cis-acting sequences.
2000,
Pubmed
,
Echinobase
Miller,
Improved retroviral vectors for gene transfer and expression.
1989,
Pubmed
Ott,
Correction of X-linked chronic granulomatous disease by gene therapy, augmented by insertional activation of MDS1-EVI1, PRDM16 or SETBP1.
2006,
Pubmed
Palla,
Enhancer blocking activity located near the 3' end of the sea urchin early H2A histone gene.
1997,
Pubmed
,
Echinobase
Pannell,
Silencing of gene expression: implications for design of retrovirus vectors.
2001,
Pubmed
Ramezani,
Performance- and safety-enhanced lentiviral vectors containing the human interferon-beta scaffold attachment region and the chicken beta-globin insulator.
2003,
Pubmed
Recillas-Targa,
Position-effect protection and enhancer blocking by the chicken beta-globin insulator are separable activities.
2002,
Pubmed
Rivella,
The cHS4 insulator increases the probability of retroviral expression at random chromosomal integration sites.
2000,
Pubmed
Roh,
The genomic landscape of histone modifications in human T cells.
2006,
Pubmed
Ryu,
A chromatin insulator blocks interactions between globin regulatory elements and cellular promoters in erythroid cells.
2007,
Pubmed
Schröder,
HIV-1 integration in the human genome favors active genes and local hotspots.
2002,
Pubmed
Spinelli,
Evidences of two different sets of histone genes active during embryogenesis of the sea urchin Paracentrotus lividus.
1979,
Pubmed
,
Echinobase
Walters,
The chicken beta-globin 5'HS4 boundary element blocks enhancer-mediated suppression of silencing.
1999,
Pubmed
West,
Insulators: many functions, many mechanisms.
2002,
Pubmed
Wu,
Transcription start regions in the human genome are favored targets for MLV integration.
2003,
Pubmed
Yannaki,
Topological constraints governing the use of the chicken HS4 chromatin insulator in oncoretrovirus vectors.
2002,
Pubmed
von Kalle,
Lenti in red: progress in gene therapy for human hemoglobinopathies.
2004,
Pubmed
