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PMID: 15475575 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Regulation of the endogenous VEGF-A gene by exogenous designed regulatory proteins.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 101 ·No. 42 ·2004-10-19 ·Pages 15225-30

Tachikawa K, Schröder O, Frey G, Briggs SP, Sera T

Abstract

We describe a facile method to activate or repress transcription of endogenous genes in a quantitative and specific manner by treatment with designed regulatory proteins (DRPs), in which artificial transcription factors (ATFs) are fused to cell-penetrating peptides (CPPs). Penetration of DRPs into cells is mediated by an N-terminal CPP fused to a nuclear localization signal; a DNA-binding domain and a transactivation domain follow. The DNA-binding domain was targeted to the vascular endothelial growth factor (VEGF)-A gene. An agonist DRP was rapidly taken up by cells and transported to the nucleus; soon after, the cells began transcribing the gene and secreting VEGF-A protein in a dose-dependent manner. Multiple copies of a short oligopeptide derived from a minimal transactivation domain of human beta-catenin was stronger than VP-16. The SRDX domain from the plant transcription factor, SUPERMAN, changed the DRP to a hypoxia-induced antagonist of VEGF-A. DRPs combine many of the potential benefits of transgenes with those of recombinant proteins.

MeSH Terms
Base Sequence Cell Line DNA/genetics Drug Design Gene Expression Regulation/drug effects Genes, Reporter Humans Protein Engineering RNA, Messenger/biosynthesis,genetics Recombinant Proteins/genetics,metabolism,pharmacology Transcription Factors/genetics,metabolism Vascular Endothelial Growth Factor A/biosynthesis,genetics
Chemicals
RNA, Messenger Recombinant Proteins Transcription Factors VEGFA protein, human Vascular Endothelial Growth Factor A DNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Tachikawa Kiyoshi
Torrey Mesa Research Institute, 3115 Merryfield Row, San Diego, CA 92121, USA. [email protected]
Schröder Oliver
Frey Gerhard
Briggs Steven P
Sera Takashi
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.0406473101
Email
[email protected]
Published
2004-10-19
Epub
2004-00-08
Pages
15225-30
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC523457
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.0406473101 PMC Full Text View journal details
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