| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
J. Biol. Chem., Vol. 275, Issue 42, 32617-32627, October 20, 2000
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
From The Skaggs Institute for Chemical Biology and the Department
of Molecular Biology, The Scripps Research Institute,
La Jolla, California 92037
Ligand-dependent transcriptional
regulators were generated by fusion of designed
Cys2-His2 zinc finger proteins and
steroid hormone receptor ligand binding domains. To produce novel DNA binding domains, three-finger proteins binding specific 9-base pair
sequences were constructed from modular building blocks. Fusion of
these zinc finger proteins to a transcriptional activation domain and
to modified ligand binding domains derived from either the estrogen or
progesterone receptors yielded potent ligand-dependent transcriptional regulators. Together with optimized minimal promoters, these regulators provide 4-hydroxytamoxifen- or RU486-inducible expression systems with induction ratios of up to 3 orders of magnitude. These inducible expression systems are functionally independent, and each can be selectively switched on within the same
cell. The potential use of zinc finger-steroid receptor fusion proteins
for the regulation of natural promoters was also explored. A
gene-specific six-finger protein binding an 18-base pair target sequence was converted into a ligand-dependent regulator by
fusion with either two estrogen receptor ligand binding domains or one ecdysone receptor and one retinoid X receptor ligand binding domain. These single-chain receptor proteins undergo an intramolecular rearrangement, rather than intermolecular dimerization and are functional as monomers. Thus, the ability to engineer DNA binding specificities of zinc finger proteins enables the construction of
ligand-dependent transcriptional regulators with potential for the regulation of virtually any desired artificial or natural promoter. It is anticipated that the novel chemically regulated gene
switches described herein will find many applications in applied and
basic research, where the specific modulation of gene expression can be exploited.
Chemically Regulated Zinc Finger Transcription Factors*
,
*
This work was supported by National Institutes of
Health Grant CA86258, Novartis, and the Skaggs Institute for Chemical
Biology (to C. F. B.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Recipient of fellowships from the Krebsliga beider Basel and the
Swiss Cancer League.
§
Present address: Novartis Pharma AG, CH-4002 Basel, Switzerland.
¶
Recipient of a fellowship from the Deutsche Forschungsgemeinschaft.
To whom correspondence should be addressed: The Scripps
Research Institute, Department of Molecular Biology, BCC-550, 10550 N. Torrey Pines Rd., La Jolla, CA 92037. Tel.: 858-784-9098; Fax: 858-784-2583; E-mail: carlos@scripps.edu.
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  B. Dreier, R. P. Fuller, D. J. Segal, C. V. Lund, P. Blancafort, A. Huber, B. Koksch, and C. F. Barbas III Development of Zinc Finger Domains for Recognition of the 5'-CNN-3' Family DNA Sequences and Their Use in the Construction of Artificial Transcription Factors J. Biol. Chem., October 21, 2005; 280(42): 35588 - 35597. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Malphettes, C. C. Weber, M. D. El-Baba, R. G. Schoenmakers, D. Aubel, W. Weber, and M. Fussenegger A novel mammalian expression system derived from components coordinating nicotine degradation in arthrobacter nicotinovorans pAO1 Nucleic Acids Res., July 7, 2005; 33(12): e107 - e107. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. J. Segal, J. Goncalves, S. Eberhardy, C. H. Swan, B. E. Torbett, X. Li, and C. F. Barbas III Attenuation of HIV-1 Replication in Primary Human Cells with a Designed Zinc Finger Transcription Factor J. Biol. Chem., April 9, 2004; 279(15): 14509 - 14519. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. G. Uil, H. J. Haisma, and M. G. Rots Therapeutic modulation of endogenous gene function by agents with designed DNA-sequence specificities Nucleic Acids Res., November 1, 2003; 31(21): 6064 - 6078. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. Guan, J. Stege, M. Kim, Z. Dahmani, N. Fan, P. Heifetz, C. F. Barbas III, and S. P. Briggs Heritable endogenous gene regulation in plants with designed polydactyl zinc finger transcription factors PNAS, October 1, 2002; 99(20): 13296 - 13301. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 R.L. Juliano, A. Astriab-Fisher, and D. Falke Macromolecular Therapeutics: Emerging Strategies for Drug Discovery in the Postgenome Era Mol. Interv., April 1, 2001; 1(1): 40 - 53. [Abstract] [Full Text]
|
|
|
|
|
|
B. Dreier, R. R. Beerli, D. J. Segal, J. D. Flippin, and C. F. Barbas III Development of Zinc Finger Domains for Recognition of the 5'-ANN-3' Family of DNA Sequences and Their Use in the Construction of Artificial Transcription Factors J. Biol. Chem., July 27, 2001; 276(31): 29466 - 29478. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
