title: Regulation of an endogenous locus using a panel of designed zinc finger proteins targeted to accessible chromatin regions. Activation of vascular endothelial growth factor A.
abstract: We have mapped conserved regions of enhanced DNase I accessibility within the endogenous chromosomal locus of vascular endothelial growth factor A (VEGF-A). Synthetic zinc finger protein (ZFP) transcription factors were designed to target DNA sequences contained within the DNase I-hypersensitive regions. These ZFPs, when fused to either VP16 or p65 transcriptional activation domains, were able to activate expression of the VEGF-A gene as assayed by mRNA accumulation and VEGF-A protein secretion through a range exceeding that induced by hypoxic stress. Importantly, multiple splice variants of VEGF-A mRNA with defined physiological functions were induced by a single engineered ZFP transcription factor. We present evidence for an enhanced activation of VEGF-A gene transcription by ZFP transcription factors fused to VP16 and p65 targeted to two distinct chromosomal sites >500 base pairs upstream or downstream of the transcription start site. Our strategy provides a novel approach for dissecting the requirements for gene regulation at a distance without altering the DNA sequence of the endogenous target locus.
authors.affiliation: Sangamo BioSciences Incorporated, Richmond, California 94804, USA.
authors.forename: P Q
authors.initials: PQ
authors.lastname: Liu
authors.forename: E J
authors.initials: EJ
authors.lastname: Rebar
authors.forename: L
authors.initials: L
authors.lastname: Zhang
authors.forename: Q
authors.initials: Q
authors.lastname: Liu
authors.forename: A C
authors.initials: AC
authors.lastname: Jamieson
authors.forename: Y
authors.initials: Y
authors.lastname: Liang
authors.forename: H
authors.initials: H
authors.lastname: Qi
authors.forename: P X
authors.initials: PX
authors.lastname: Li
authors.forename: B
authors.initials: B
authors.lastname: Chen
authors.forename: M C
authors.initials: MC
authors.lastname: Mendel
authors.forename: X
authors.initials: X
authors.lastname: Zhong
authors.forename: Y L
authors.initials: YL
authors.lastname: Lee
authors.forename: S P
authors.initials: SP
authors.lastname: Eisenberg
authors.forename: S K
authors.initials: SK
authors.lastname: Spratt
authors.forename: C C
authors.initials: CC
authors.lastname: Case
authors.forename: A P
authors.initials: AP
authors.lastname: Wolffe
mesh_terms: D002461:Cell Line, Transformed; D002874:Chromosome Mapping; D003850:Deoxyribonuclease I / Q000235:genetics*; D016228:Endothelial Growth Factors / Q000235:genetics*; D006801:Humans; D011506:Proteins / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D015533:Transcriptional Activation; D042461:Vascular Endothelial Growth Factor A; D016335:Zinc Fingers
chemical_list: D016228:Endothelial Growth Factors; D011506:Proteins; D014157:Transcription Factors; D042461:Vascular Endothelial Growth Factor A; D003850:Deoxyribonuclease I