Inhibition of human immunodeficiency virus type 1 replication with artificial transcription factors targeting the highly conserved primer-binding site.
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.
Eberhardy
Scott R
SR
The Skaggs Institute for Chemical Biology, La Jolla, California 92037, USA.
Goncalves
Joao
J
Coelho
Sofia
S
Segal
David J
DJ
Berkhout
Ben
B
Barbas
Carlos F
CF
D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D016325:HIV Long Terminal Repeat / Q000502:physiology; D015497:HIV-1 / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D014158:Transcription, Genetic; D014779:Virus Replication / Q000187:drug effects*; D016335:Zinc Fingers / Q000235:genetics*
D012346:RNA, Transfer, Amino Acyl; D014157:Transcription Factors