Synthetic zinc finger transcription factor action at an endogenous chromosomal site

Activation of the human erythropoietin gene

Lei Zhang, S. Kaye Spratt, Qiang Liu, Brian Johnstone, Hong Qi, Eva E. Raschke, Andrew C. Jamieson, Edward J. Rebar, Alan P. Wolffe, Casey C. Case

Research output: Contribution to journalArticle

151 Citations (Scopus)

Abstract

We have targeted the activation of an endogenous chromosomal locus including the human erythropoietin gene using synthetic transcription factors. These transcription factors are targeted to particular DNA sequences in the 5'-flanking region of the erythropoietin gene through engineering of a zinc finger DNA binding domain. The DNA binding domain is linked to a VP16 transcriptional activation domain. We find that these synthetic transcription factors invariably activate transiently transfected templates in which sequences within the 5' flank of the erythropoietin gene are fused to a luciferase reporter. The efficiency of activation under these circumstances at a defined site is dependent on DNA binding affinity. In contrast, only a subset of these same zinc finger proteins is able to activate the endogenous chromosomal locus. The activity of these proteins is influenced by their capacity to gain access to their recognition elements within the chromatin infrastructure. Zinc finger transcription factors will provide a powerful tool to probe the determinants of chromatin accessibility and remodeling within endogenous chromosomal loci.

Original languageEnglish (US)
Pages (from-to)33850-33860
Number of pages11
JournalJournal of Biological Chemistry
Volume275
Issue number43
DOIs
StatePublished - Oct 27 2000
Externally publishedYes

Fingerprint

Zinc Fingers
Erythropoietin
Zinc
Transcription Factors
Genes
Chemical activation
Chromatin
DNA
Synthetic Genes
Chromatin Assembly and Disassembly
5' Flanking Region
DNA sequences
Luciferases
Transcriptional Activation
Proteins

ASJC Scopus subject areas

  • Biochemistry

Cite this

Synthetic zinc finger transcription factor action at an endogenous chromosomal site : Activation of the human erythropoietin gene. / Zhang, Lei; Spratt, S. Kaye; Liu, Qiang; Johnstone, Brian; Qi, Hong; Raschke, Eva E.; Jamieson, Andrew C.; Rebar, Edward J.; Wolffe, Alan P.; Case, Casey C.

In: Journal of Biological Chemistry, Vol. 275, No. 43, 27.10.2000, p. 33850-33860.

Research output: Contribution to journalArticle

Zhang, L, Spratt, SK, Liu, Q, Johnstone, B, Qi, H, Raschke, EE, Jamieson, AC, Rebar, EJ, Wolffe, AP & Case, CC 2000, 'Synthetic zinc finger transcription factor action at an endogenous chromosomal site: Activation of the human erythropoietin gene', Journal of Biological Chemistry, vol. 275, no. 43, pp. 33850-33860. https://doi.org/10.1074/jbc.M005341200
Zhang, Lei ; Spratt, S. Kaye ; Liu, Qiang ; Johnstone, Brian ; Qi, Hong ; Raschke, Eva E. ; Jamieson, Andrew C. ; Rebar, Edward J. ; Wolffe, Alan P. ; Case, Casey C. / Synthetic zinc finger transcription factor action at an endogenous chromosomal site : Activation of the human erythropoietin gene. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 43. pp. 33850-33860.
@article{1e58dc1c7e37470eb19269e28d8fd54d,
title = "Synthetic zinc finger transcription factor action at an endogenous chromosomal site: Activation of the human erythropoietin gene",
abstract = "We have targeted the activation of an endogenous chromosomal locus including the human erythropoietin gene using synthetic transcription factors. These transcription factors are targeted to particular DNA sequences in the 5'-flanking region of the erythropoietin gene through engineering of a zinc finger DNA binding domain. The DNA binding domain is linked to a VP16 transcriptional activation domain. We find that these synthetic transcription factors invariably activate transiently transfected templates in which sequences within the 5' flank of the erythropoietin gene are fused to a luciferase reporter. The efficiency of activation under these circumstances at a defined site is dependent on DNA binding affinity. In contrast, only a subset of these same zinc finger proteins is able to activate the endogenous chromosomal locus. The activity of these proteins is influenced by their capacity to gain access to their recognition elements within the chromatin infrastructure. Zinc finger transcription factors will provide a powerful tool to probe the determinants of chromatin accessibility and remodeling within endogenous chromosomal loci.",
author = "Lei Zhang and Spratt, {S. Kaye} and Qiang Liu and Brian Johnstone and Hong Qi and Raschke, {Eva E.} and Jamieson, {Andrew C.} and Rebar, {Edward J.} and Wolffe, {Alan P.} and Case, {Casey C.}",
year = "2000",
month = "10",
day = "27",
doi = "10.1074/jbc.M005341200",
language = "English (US)",
volume = "275",
pages = "33850--33860",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "43",

}

TY - JOUR

T1 - Synthetic zinc finger transcription factor action at an endogenous chromosomal site

T2 - Activation of the human erythropoietin gene

AU - Zhang, Lei

AU - Spratt, S. Kaye

AU - Liu, Qiang

AU - Johnstone, Brian

AU - Qi, Hong

AU - Raschke, Eva E.

AU - Jamieson, Andrew C.

AU - Rebar, Edward J.

AU - Wolffe, Alan P.

AU - Case, Casey C.

PY - 2000/10/27

Y1 - 2000/10/27

N2 - We have targeted the activation of an endogenous chromosomal locus including the human erythropoietin gene using synthetic transcription factors. These transcription factors are targeted to particular DNA sequences in the 5'-flanking region of the erythropoietin gene through engineering of a zinc finger DNA binding domain. The DNA binding domain is linked to a VP16 transcriptional activation domain. We find that these synthetic transcription factors invariably activate transiently transfected templates in which sequences within the 5' flank of the erythropoietin gene are fused to a luciferase reporter. The efficiency of activation under these circumstances at a defined site is dependent on DNA binding affinity. In contrast, only a subset of these same zinc finger proteins is able to activate the endogenous chromosomal locus. The activity of these proteins is influenced by their capacity to gain access to their recognition elements within the chromatin infrastructure. Zinc finger transcription factors will provide a powerful tool to probe the determinants of chromatin accessibility and remodeling within endogenous chromosomal loci.

AB - We have targeted the activation of an endogenous chromosomal locus including the human erythropoietin gene using synthetic transcription factors. These transcription factors are targeted to particular DNA sequences in the 5'-flanking region of the erythropoietin gene through engineering of a zinc finger DNA binding domain. The DNA binding domain is linked to a VP16 transcriptional activation domain. We find that these synthetic transcription factors invariably activate transiently transfected templates in which sequences within the 5' flank of the erythropoietin gene are fused to a luciferase reporter. The efficiency of activation under these circumstances at a defined site is dependent on DNA binding affinity. In contrast, only a subset of these same zinc finger proteins is able to activate the endogenous chromosomal locus. The activity of these proteins is influenced by their capacity to gain access to their recognition elements within the chromatin infrastructure. Zinc finger transcription factors will provide a powerful tool to probe the determinants of chromatin accessibility and remodeling within endogenous chromosomal loci.

UR - http://www.scopus.com/inward/record.url?scp=0034721877&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034721877&partnerID=8YFLogxK

U2 - 10.1074/jbc.M005341200

DO - 10.1074/jbc.M005341200

M3 - Article

VL - 275

SP - 33850

EP - 33860

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 43

ER -