RRC ID 55984
Author Riani YD, Matsuda T, Takemoto K, Nagai T.
Title Green monomeric photosensitizing fluorescent protein for photo-inducible protein inactivation and cell ablation.
Journal BMC Biol
Abstract BACKGROUND:Photosensitizing fluorescent proteins, which generate reactive oxygen species (ROS) upon light irradiation, are useful for spatiotemporal protein inactivation and cell ablation. They give us clues about protein function, intracellular signaling pathways and intercellular interactions. Since ROS generation of a photosensitizer is specifically controlled by certain excitation wavelengths, utilizing colour variants of photosensitizing protein would allow multi-spatiotemporal control of inactivation. To expand the colour palette of photosensitizing protein, here we developed SuperNova Green from its red predecessor, SuperNova.
RESULTS:SuperNova Green is able to produce ROS spatiotemporally upon blue light irradiation. Based on protein characterization, SuperNova Green produces insignificant amounts of singlet oxygen and predominantly produces superoxide and its derivatives. We utilized SuperNova Green to specifically inactivate the pleckstrin homology domain of phospholipase C-δ1 and to ablate cancer cells in vitro. As a proof of concept for multi-spatiotemporal control of inactivation, we demonstrate that SuperNova Green can be used with its red variant, SuperNova, to perform independent protein inactivation or cell ablation studies in a spatiotemporal manner by selective light irradiation.
CONCLUSION:Development of SuperNova Green has expanded the photosensitizing protein toolbox to optogenetically control protein inactivation and cell ablation.
Volume 16(1)
Pages 50
Published 2018-4-30
DOI 10.1186/s12915-018-0514-7
PII 10.1186/s12915-018-0514-7
PMID 29712573
PMC PMC5928576
MeSH Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism* Light* Photosensitizing Agents Reactive Oxygen Species / metabolism Singlet Oxygen / metabolism Superoxides / metabolism
IF 6.723
Times Cited 5
Resource
Human and Animal Cells HeLa