Document Detail


Detection of apoptosis using cyclic luciferase in living mammals.
MedLine Citation:
PMID:  19685303     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Programmed cell death, apoptosis, is a crucial process involved in pathogenesis and progression of diseases, which is executed by cysteine aspartyl proteases (caspases). The caspase activities in living subjects and their regulation with small chemical compounds are of great interest for screening drug candidates or pathological agents. We describe a novel genetically encoded bioluminescent indicator for real-time imaging of caspase-3 activities in living cells and animals. The indicator is composed of an engineered firefly luciferase, of which the N- and C-terminal ends are linked with a substrate sequence of caspase-3 (Asp-Glu-Val-Asp). When activated caspase-3 digests the substrate sequence, the cyclized luciferase recovers its activity. The indicator provides a general means of evaluating effects of cytotoxic compounds or novel pharmacological chemicals and apoptosis.
Authors:
Akira Kanno; Yoshio Umezawa; Takeaki Ozawa
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Methods in molecular biology (Clifton, N.J.)     Volume:  574     ISSN:  1940-6029     ISO Abbreviation:  Methods Mol. Biol.     Publication Date:  2009  
Date Detail:
Created Date:  2009-08-17     Completed Date:  2010-01-11     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9214969     Medline TA:  Methods Mol Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  105-14     Citation Subset:  IM    
Affiliation:
Department of Chemistry, School of Science, The University of Tokyo, Tokyo, Japan.
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MeSH Terms
Descriptor/Qualifier:
Animals
Apoptosis*
Base Sequence
Caspase 3 / metabolism
DNA Primers
Hela Cells
Humans
Luciferases / genetics*
Plasmids
Polymerase Chain Reaction
Chemical
Reg. No./Substance:
0/DNA Primers; EC 1.13.12.-/Luciferases; EC 3.4.22.-/Caspase 3

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