The GATA1-HS2 enhancer allows persistent and position-independent expression of a β-globin transgene

Annarita Miccio, Valentina Poletti, Francesca Tiboni, Claudia Rossi, Antonella Antonelli, Fulvio Mavilio, Giuliana Ferrari

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Gene therapy of genetic diseases requires persistent and position-independent expression of a therapeutic transgene. Transcriptional enhancers binding chromatin-remodeling and modifying complexes may play a role in shielding transgenes from repressive chromatin effects. We tested the activity of the HS2 enhancer of the GATA1 gene in protecting the expression of a β-globin minigene delivered by a lentiviral vector in hematopoietic stem/progenitor cells. Gene expression from proviruses carrying GATA1-HS2 in both LTRs was persistent and resistant to silencing at most integration sites in the in vivo progeny of human hematopoietic progenitors and murine long-term repopulating stem cells. The GATA1-HS2-modified vector allowed correction of murine β-thalassemia at low copy number without inducing clonal selection of erythroblastic progenitors. Chromatin immunoprecipitation studies showed that GATA1 and the CBP acetyltransferase bind to GATA1-HS2, significantly increasing CBP-specific histone acetylations at the LTRs and β-globin promoter. Recruitment of CBP by the LTRs thus establishes an open chromatin domain encompassing the entire provirus, and increases the therapeutic efficacy of β-globin gene transfer by reducing expression variegation and epigenetic silencing.

Original languageEnglish
Article numbere27955
JournalPLoS One
Volume6
Issue number12
DOIs
Publication statusPublished - Dec 2 2011

Fingerprint

Globins
Transgenes
transgenes
Chromatin
chromatin
Proviruses
Hematopoietic Stem Cells
proviruses
stem cells
Stem cells
Inborn Genetic Diseases
Acetyltransferases
Chromatin Assembly and Disassembly
Thalassemia
Chromatin Immunoprecipitation
Acetylation
thalassemia
Gene transfer
Epigenomics
Gene therapy

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

The GATA1-HS2 enhancer allows persistent and position-independent expression of a β-globin transgene. / Miccio, Annarita; Poletti, Valentina; Tiboni, Francesca; Rossi, Claudia; Antonelli, Antonella; Mavilio, Fulvio; Ferrari, Giuliana.

In: PLoS One, Vol. 6, No. 12, e27955, 02.12.2011.

Research output: Contribution to journalArticle

Miccio, Annarita ; Poletti, Valentina ; Tiboni, Francesca ; Rossi, Claudia ; Antonelli, Antonella ; Mavilio, Fulvio ; Ferrari, Giuliana. / The GATA1-HS2 enhancer allows persistent and position-independent expression of a β-globin transgene. In: PLoS One. 2011 ; Vol. 6, No. 12.
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