Attenuation of HIV-1 Replication in Primary Human Cells with a Designed Zinc Finger Transcription Factor

David Segal, João Gonçalves, Scott Eberhardy, Christina H. Swan, Bruce E. Torbett, Xuelin Li, Carlos F. Barbas

Research output: Contribution to journalArticle

68 Citations (Scopus)

Abstract

Small molecule inhibitors of human immunodeficiency virus, type 1 (HIV-1) have been extremely successful but are associated with a myriad of undesirable effects and require lifelong daily dosing. In this study we explore an alternative approach, that of inducing intracellular immunity using designed, zinc finger-based transcription factors. Three transcriptional repression proteins were engineered to bind sites in the HIV-1 promoter that were expected to be both accessible in chromatin structure and highly conserved in sequence structure among the various HIV-1 subgroups. Transient transfection assays identified one factor, KRAB-HLTR3, as being able to achieve 100-fold repression of an HIV-1 promoter. Specificity of repression was demonstrated by the lack of repression of other promoters. This factor was further shown to repress the replication of several HIV-1 viral strains 10- to 100-fold in T-cell lines and primary human peripheral blood mononuclear cells. Repression was observed for at least 18 days with no significant cytotoxicity. Stable T-cell lines expressing the factor also do not show obvious signs of cytotoxicity. These characteristics present KRAB-HLTR3 as an attractive candidate for development in an intracellular immunization strategy for anti-HIV-1 therapy.

Original languageEnglish (US)
Pages (from-to)14509-14519
Number of pages11
JournalJournal of Biological Chemistry
Volume279
Issue number15
DOIs
StatePublished - Apr 9 2004
Externally publishedYes

Fingerprint

Zinc Fingers
Virus Replication
Viruses
HIV-1
Zinc
Transcription Factors
Cells
T-cells
Cytotoxicity
Immunization
T-Lymphocytes
Cell Line
Conserved Sequence
Chromatin
Transfection
Immunity
Assays
Blood Cells
Blood
Molecules

ASJC Scopus subject areas

  • Biochemistry

Cite this

Attenuation of HIV-1 Replication in Primary Human Cells with a Designed Zinc Finger Transcription Factor. / Segal, David; Gonçalves, João; Eberhardy, Scott; Swan, Christina H.; Torbett, Bruce E.; Li, Xuelin; Barbas, Carlos F.

In: Journal of Biological Chemistry, Vol. 279, No. 15, 09.04.2004, p. 14509-14519.

Research output: Contribution to journalArticle

Segal, David ; Gonçalves, João ; Eberhardy, Scott ; Swan, Christina H. ; Torbett, Bruce E. ; Li, Xuelin ; Barbas, Carlos F. / Attenuation of HIV-1 Replication in Primary Human Cells with a Designed Zinc Finger Transcription Factor. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 15. pp. 14509-14519.
@article{a1e10312c4a24b57808a169e6a4d9636,
title = "Attenuation of HIV-1 Replication in Primary Human Cells with a Designed Zinc Finger Transcription Factor",
abstract = "Small molecule inhibitors of human immunodeficiency virus, type 1 (HIV-1) have been extremely successful but are associated with a myriad of undesirable effects and require lifelong daily dosing. In this study we explore an alternative approach, that of inducing intracellular immunity using designed, zinc finger-based transcription factors. Three transcriptional repression proteins were engineered to bind sites in the HIV-1 promoter that were expected to be both accessible in chromatin structure and highly conserved in sequence structure among the various HIV-1 subgroups. Transient transfection assays identified one factor, KRAB-HLTR3, as being able to achieve 100-fold repression of an HIV-1 promoter. Specificity of repression was demonstrated by the lack of repression of other promoters. This factor was further shown to repress the replication of several HIV-1 viral strains 10- to 100-fold in T-cell lines and primary human peripheral blood mononuclear cells. Repression was observed for at least 18 days with no significant cytotoxicity. Stable T-cell lines expressing the factor also do not show obvious signs of cytotoxicity. These characteristics present KRAB-HLTR3 as an attractive candidate for development in an intracellular immunization strategy for anti-HIV-1 therapy.",
author = "David Segal and Jo{\~a}o Gon{\cc}alves and Scott Eberhardy and Swan, {Christina H.} and Torbett, {Bruce E.} and Xuelin Li and Barbas, {Carlos F.}",
year = "2004",
month = "4",
day = "9",
doi = "10.1074/jbc.M400349200",
language = "English (US)",
volume = "279",
pages = "14509--14519",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "15",

}

TY - JOUR

T1 - Attenuation of HIV-1 Replication in Primary Human Cells with a Designed Zinc Finger Transcription Factor

AU - Segal, David

AU - Gonçalves, João

AU - Eberhardy, Scott

AU - Swan, Christina H.

AU - Torbett, Bruce E.

AU - Li, Xuelin

AU - Barbas, Carlos F.

PY - 2004/4/9

Y1 - 2004/4/9

N2 - Small molecule inhibitors of human immunodeficiency virus, type 1 (HIV-1) have been extremely successful but are associated with a myriad of undesirable effects and require lifelong daily dosing. In this study we explore an alternative approach, that of inducing intracellular immunity using designed, zinc finger-based transcription factors. Three transcriptional repression proteins were engineered to bind sites in the HIV-1 promoter that were expected to be both accessible in chromatin structure and highly conserved in sequence structure among the various HIV-1 subgroups. Transient transfection assays identified one factor, KRAB-HLTR3, as being able to achieve 100-fold repression of an HIV-1 promoter. Specificity of repression was demonstrated by the lack of repression of other promoters. This factor was further shown to repress the replication of several HIV-1 viral strains 10- to 100-fold in T-cell lines and primary human peripheral blood mononuclear cells. Repression was observed for at least 18 days with no significant cytotoxicity. Stable T-cell lines expressing the factor also do not show obvious signs of cytotoxicity. These characteristics present KRAB-HLTR3 as an attractive candidate for development in an intracellular immunization strategy for anti-HIV-1 therapy.

AB - Small molecule inhibitors of human immunodeficiency virus, type 1 (HIV-1) have been extremely successful but are associated with a myriad of undesirable effects and require lifelong daily dosing. In this study we explore an alternative approach, that of inducing intracellular immunity using designed, zinc finger-based transcription factors. Three transcriptional repression proteins were engineered to bind sites in the HIV-1 promoter that were expected to be both accessible in chromatin structure and highly conserved in sequence structure among the various HIV-1 subgroups. Transient transfection assays identified one factor, KRAB-HLTR3, as being able to achieve 100-fold repression of an HIV-1 promoter. Specificity of repression was demonstrated by the lack of repression of other promoters. This factor was further shown to repress the replication of several HIV-1 viral strains 10- to 100-fold in T-cell lines and primary human peripheral blood mononuclear cells. Repression was observed for at least 18 days with no significant cytotoxicity. Stable T-cell lines expressing the factor also do not show obvious signs of cytotoxicity. These characteristics present KRAB-HLTR3 as an attractive candidate for development in an intracellular immunization strategy for anti-HIV-1 therapy.

UR - http://www.scopus.com/inward/record.url?scp=2442536906&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2442536906&partnerID=8YFLogxK

U2 - 10.1074/jbc.M400349200

DO - 10.1074/jbc.M400349200

M3 - Article

VL - 279

SP - 14509

EP - 14519

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 15

ER -