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PMID: 12219076 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

A light-switchable gene promoter system.

Nature biotechnology ·Vol. 20 ·No. 10 ·2002-10-00 ·Pages 1041-4

Shimizu-Sato S, Huq E, Tepperman JM, Quail PH

Abstract

Regulatable transgene systems providing easily controlled, conditional induction or repression of expression are indispensable tools in biomedical and agricultural research and biotechnology. Several such systems have been developed for eukaryotes. Most of these rely on the administration of either exogenous chemicals or heat shock. Despite the general success of many of these systems, the potential for problems, such as toxic, unintended, or pleiotropic effects of the inducing chemical or treatment, can impose limitations on their use. We have developed a promoter system that can be induced, rapidly and reversibly, by short pulses of light. This system is based on the known red light-induced binding of the plant photoreceptor phytochrome to the protein PIF3 and the reversal of this binding by far-red light. We show here that yeast cells expressing two chimeric proteins, a phytochrome-GAL4-DNA-binding-domain fusion and a PIF3-GAL4-activation-domain fusion, are induced by red light to express selectable or "scorable" marker genes containing promoters with a GAL4 DNA-binding site, and that this induction is rapidly abrogated by subsequent far-red light. We further show that the extent of induction can be controlled precisely by titration of the number of photons delivered to the cells by the light pulse. Thus, this system has the potential to provide rapid, noninvasive, switchable control of the expression of a desired gene to a preselected level in any suitable cell by simple exposure to a light signal.

MeSH Terms
Arabidopsis Proteins Basic Helix-Loop-Helix Transcription Factors Cells, Cultured Dose-Response Relationship, Radiation Gene Expression Regulation, Fungal/radiation effects Gene Transfer Techniques Light Photosynthetic Reaction Center Complex Proteins/genetics,metabolism Phytochrome/genetics,metabolism Promoter Regions, Genetic Recombinant Fusion Proteins Signal Transduction/genetics,radiation effects Species Specificity Transgenes Yeasts/classification,genetics,metabolism
Chemicals
Arabidopsis Proteins Basic Helix-Loop-Helix Transcription Factors PIF3 protein, Arabidopsis Photosynthetic Reaction Center Complex Proteins Recombinant Fusion Proteins Phytochrome
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Shimizu-Sato Sae
Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720, USA.
Huq Enamul
Tepperman James M
Quail Peter H
Article Info
Journal
Nature biotechnology
Abbr.
Nat Biotechnol
ISSN
1087-0156
Published
2002-10-00
Epub
2002-00-03
Pages
1041-4
Language
English
Region
United States
NLM ID
9604648
Subset
IM
Grants
NIGMS NIH HHS · GM47475 · United States
Corrections
CommentIn
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