1. 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
- Author
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P Q, Liu, E J, Rebar, L, Zhang, Q, Liu, A C, Jamieson, Y, Liang, H, Qi, P X, Li, B, Chen, M C, Mendel, X, Zhong, Y L, Lee, S P, Eisenberg, S K, Spratt, C C, Case, and A P, Wolffe
- Subjects
Transcriptional Activation ,Vascular Endothelial Growth Factor A ,Chromosome Mapping ,Deoxyribonuclease I ,Humans ,Proteins ,Zinc Fingers ,Endothelial Growth Factors ,Cell Line, Transformed ,Transcription Factors - 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 sites500 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.
- Published
- 2001