Inhibition of human immunodeficiency virus type 1 replication with artificial transcription factors targeting the highly conserved primer-binding site

Scott R. Eberhardy, Joao Goncalves, Sofia Coelho, David Segal, Ben Berkhout, Carlos F. Barbas

Research output: Contribution to journalArticle

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Abstract

The human immunodeficiency virus type 1 (HIV-1) primer-binding site (PBS) is a highly conserved region in the HIV genome and represents an attractive target for the development of new anti-HIV therapies. In this study, we designed four artificial zinc finger transcription factors to bind at or adjacent to the PBS and repress transcription from the HIV-1 long terminal repeat (LTR). These proteins bound to the LTR in vivo, as demonstrated by the chromatin immunoprecipitation assay. In transient reporter assays, three of the four proteins repressed transcription of a reporter driven by the HIV-1 LTR. Only one of these proteins, however, designated KRAB-PBS2, was able to prevent virus production when transduced into primary lymphocytes. We observed >90% inhibition of viral replication over the course of several weeks compared to untransduced cells, and no significant cytotoxicity was observed. Long-term exposure of HIV-1 to KRAB-PBS2 induced mutations in the HIV-1 PBS that reduced the effectiveness of the repressor, but these mutations also resulted in decreased rates of viral replication. These results show that KRAB-PBS2 has the potential to be used in antiviral therapy for AIDS patients and might complement other gene-based strategies.

Original languageEnglish (US)
Pages (from-to)2873-2883
Number of pages11
JournalJournal of Virology
Volume80
Issue number6
DOIs
StatePublished - Mar 2006

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Virus Replication
Human immunodeficiency virus 1
HIV-1
binding sites
Transcription Factors
transcription factors
Binding Sites
terminal repeat sequences
HIV Long Terminal Repeat
virus replication
transcription (genetics)
HIV
therapeutics
Mutation
Proteins
proteins
Terminal Repeat Sequences
Chromatin Immunoprecipitation
zinc finger motif
Zinc Fingers

ASJC Scopus subject areas

  • Immunology

Cite this

Inhibition of human immunodeficiency virus type 1 replication with artificial transcription factors targeting the highly conserved primer-binding site. / Eberhardy, Scott R.; Goncalves, Joao; Coelho, Sofia; Segal, David; Berkhout, Ben; Barbas, Carlos F.

In: Journal of Virology, Vol. 80, No. 6, 03.2006, p. 2873-2883.

Research output: Contribution to journalArticle

Eberhardy, Scott R. ; Goncalves, Joao ; Coelho, Sofia ; Segal, David ; Berkhout, Ben ; Barbas, Carlos F. / Inhibition of human immunodeficiency virus type 1 replication with artificial transcription factors targeting the highly conserved primer-binding site. In: Journal of Virology. 2006 ; Vol. 80, No. 6. pp. 2873-2883.
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