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Deleterious effects of mitochondrial ROS generated by KillerRed photodynamic action in human cell lines and C. elegans

Paper ID Volume ID Publish Year Pages File Format Full-Text
30647 44495 2012 12 PDF Available
Title
Deleterious effects of mitochondrial ROS generated by KillerRed photodynamic action in human cell lines and C. elegans
Abstract

KillerRed, a red fluorescent protein, is a photosensitizer that efficiently generates reactive oxygen species (ROS) when irradiated with green light. Because KillerRed is genetically encoded, it can be expressed in a spatially and temporally regulated manner under control of a chosen promoter and thus is a powerful tool for studying the downstream cellular effects of ROS. However, information is still limited about the effects of KillerRed-mediated production of ROS inside the mitochondria (mtROS). Therefore, we investigated whether mtROS generated by KillerRed could trigger mitochondrial damage and cell death by generating human cell lines (HEK293T and HeLa cells) that stably expressed mitochondria-targeting KillerRed (mtKillerRed). We found that mtROS generated by mtKillerRed caused depolarization of the mitochondrial membrane and morphological changes, which were partly due to the mitochondrial permeability transition (MPT), as well as inducing both caspase-dependent cell death (apoptosis) and caspase-independent cell death. In order to study the pathological processes initiated by mtROS in animals, transgenic Caenorhabditis elegans expressing mtKillerRed in muscle tissue were generated. Transgenic larvae showed developmental delay following light irradiation, suggesting that mtROS influenced the development of C. elegans larvae. In conclusion, our studies demonstrated that the photosensitizer KillerRed is effective at inducing oxidative damage in the mitochondria, and indicated that our experimental systems may be useful for studying the downstream cellular effects of mtROS.

► mtROS generated by KillerRed could trigger mitochondrial damage. ► mtROS induced caspase-dependent and caspase-independent cell death in human cells. ► mtKillerRed alone showed toxicity in C. elegans under unirradiated conditions. ► Photodynamic action of mtKillerRed induced developmental delay of C. elegans larvae.

Keywords
KillerRed; Photosensitizer; Mitochondrial ROS; Photodynamic therapy (PDT); C. elegans
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Deleterious effects of mitochondrial ROS generated by KillerRed photodynamic action in human cell lines and C. elegans
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Photochemistry and Photobiology B: Biology - Volume 117, 5 December 2012, Pages 1–12
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us