☆ 4.6 Article

Rational design of minimal hypoxia-inducible enhancers

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS (2008)

期刊

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS

卷 370, 期 4, 页码 613-618

出版社

ACADEMIC PRESS INC ELSEVIER SCIENCE

DOI: 10.1016/j.bbrc.2008.03.147

关键词

hypoxia; hypoxia-inducible factor; hypoxia responsive element; HIF-binding site

类别

Biochemistry & Molecular Biology Biophysics

资金

NCI NIH HHS [R01 CA104214-03S1] Funding Source: Medline

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Protocol

Reagent

摘要

The hypoxia-inducible factor (HIF) activates transcription via binding to the highly variable hypoxia-responsive elements (HREs). All hypoxia-inducible constructs described to date utilize multimers of naturally occurring HREs. Here, we describe the rational design of minimal hypoxia-inducible enhancers, conceptually equivalent to using an optimized HIF-binding site (HBS) as the building block. Optimizations of the HBS, spacing between HBSs, the distance from the minimal promoter, and orientation of HBSs allowed us to design constructs with high hypoxic activity. Activation of the 4xopt HBS (36 bp) construct by hypoxia or HIF-1 alpha and HIF-2 alpha was comparable with that of the 4xEPO HRE (208 bp) construct. The strong synergism between the properly arranged optimized HBSs was due to stimulation of high affinity HIF binding. Our data prove, for the first time, that it is possible to assemble artificial hypoxia-inducible enhancers from a single type of regulatory element-optimized HBS. (c) 2008 Elsevier Inc. All rights reserved.

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