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Detection of Apoptosis Using Cyclic Luciferase in Living Mammals

  • Akira Kanno
  • Yoshio Umezawa
  • Takeaki Ozawa
Part of the Methods in Molecular Biology™ book series (MIMB, volume 574)

Abstract

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.

Key words

Apoptosis caspases imaging luciferases luminescence 

Notes

Acknowledgment

This work was supported by grants from the Japan Science and Technology Agency (JST), and Japan Society for the Promotion of Science (JSPS).

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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Akira Kanno
    • 1
  • Yoshio Umezawa
    • 2
  • Takeaki Ozawa
    • 1
  1. 1.Department of Chemistry, School of ScienceThe University of Tokyo; Japan Science and Technology AgencyTokyoJapan
  2. 2.Department of Chemistry, School of ScienceThe University of Tokyo; Research Institute of Pharmaceutical SciencesTokyoJapan

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