Document Detail


Photoactivated gene expression for cell fate mapping and cell manipulation.
MedLine Citation:
PMID:  11752627     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A long-standing goal of developmental biologists is to create developmental fate maps by tracking individual cells through development. Another objective is to perturb the behavior of selected cells and follow the ensuing effects. To this end, we have developed a technique that allows for spatial and temporal control of gene expression in single cells or patches of cells using light to induce gene expression. This technique relies on "caging" the activity of the potent transcriptional activator GAL4VP16 with a photolabile compound, which can be removed with a brief exposure to long-wavelength ultraviolet (UV) light. The caged GAL4VP16 is injected into early-stage embryos, which are aged to the desired point in development, and the cell(s) of interest are irradiated with a brief pulse of long-wavelength UV light. This method has been used extensively in Drosophila, Xenopus, and Zebrafish embryos. The methods for purifying, caging, injection, and photoactivation of the GAL4VP16 protein, and methods for the visualization of marked cells are described in detail.
Authors:
J Minden; R Namba; J Mergliano; S Cambridge
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Publication Detail:
Type:  Journal Article     Date:  2000-12-12
Journal Detail:
Title:  Science's STKE : signal transduction knowledge environment     Volume:  2000     ISSN:  1525-8882     ISO Abbreviation:  Sci. STKE     Publication Date:  2000 Dec 
Date Detail:
Created Date:  2001-12-25     Completed Date:  2002-02-07     Revised Date:  2009-10-21    
Medline Journal Info:
Nlm Unique ID:  100964423     Medline TA:  Sci STKE     Country:  United States    
Other Details:
Languages:  eng     Pagination:  pl1     Citation Subset:  IM    
Affiliation:
Department of Biological Sciences and Center for Light Microscope Imaging and Biotechnology, Carnegie Mellon University, Pittsburgh, PA, USA. minden@cmu.edu
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MeSH Terms
Descriptor/Qualifier:
Animals
Cell Lineage / genetics,  radiation effects*
Drosophila melanogaster / embryology,  genetics,  radiation effects
Embryo, Nonmammalian / chemistry,  cytology
Gene Expression Regulation, Developmental / genetics,  radiation effects*
Genes, Insect / genetics,  radiation effects
Photochemistry
Xenopus laevis / embryology,  genetics
Zebrafish / embryology,  genetics

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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