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Human immunodeficiency virus type 1 incorporated with fusion proteins consisting of integrase and the designed polydactyl zinc finger protein e2c can bias integration of viral DNA into a predetermined chromosomal region in human cells

Academic Article
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Overview

authors

  • Tan, W. J.
  • Dong, Z.
  • Wilkinson, T. A.
  • Barbas III, Carlos
  • Chow, S. A.

publication date

  • February 2006

journal

  • Journal of Virology  Journal

abstract

  • In vitro studies using fusion proteins consisting of human immunodeficiency virus type 1 integrase (IN) and a synthetic polydactyl zinc finger protein E2C, a sequence-specific DNA-binding protein, showed that integration of retroviral DNA can be biased towards a contiguous 18-bp E2C-recognition site. To determine whether the fusion protein strategy can achieve site-specific integration in vivo, viruses were prepared by cotransfection and various IN-E2C fusion proteins were packaged in trans into virions. The resulting viruses incorporated with the IN-E2C fusion proteins were functional and capable of performing integration at a level ranging from 1 to 24% of that of viruses containing wild-type (WT) IN. Two of the more infectious viruses, which contained E2C fused to either the N (E2C/IN) or to the C (IN/E2C) terminus of IN, were tested for their ability to direct integration into a unique E2C-binding site present within the 5' untranslated region of erbB-2 gene on human chromosome 17. The copy number of proviral DNA was measured using a quantitative real-time nested-PCR assay, and the specificity of directed integration was determined by comparing the number of proviruses within the vicinity of the E2C-binding site to that in the whole genome. Viruses containing IN/E2C fusion proteins had sevenfold higher preference for integrating near the E2C-binding site than those viruses containing WT IN, whereas viruses containing E2C/IN had 10-fold higher preference. The results indicated that the IN-E2C fusion protein strategy is capable of directing integration of retroviral DNA into a predetermined chromosomal region in the human genome.

subject areas

  • Binding Sites
  • Cell Line
  • Chromosomes, Human, Pair 17
  • DNA, Viral
  • DNA-Binding Proteins
  • Genes, erbB-2
  • Genome, Human
  • HIV Integrase
  • HIV-1
  • Humans
  • Polymerase Chain Reaction
  • Proviruses
  • Recombinant Fusion Proteins
  • Virus Integration
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Identity

International Standard Serial Number (ISSN)

  • 0022-538X

Digital Object Identifier (DOI)

  • 10.1128/jvi.80.4.1939-1948.2006

PubMed ID

  • 16439549
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Additional Document Info

start page

  • 1939

end page

  • 1948

volume

  • 80

issue

  • 4

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