Source 1review2014Journal of Cell Science
Toolkit/KillerRed
KillerRed
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
KillerRed is a genetically encoded protein photosensitizer used to generate singlet oxygen upon light irradiation for photodynamic perturbation. In the cited study, plasma-membrane-targeted KillerRed activated cholecystokinin 1 receptor (CCK1R) signaling in AR4-2J cells, producing persistent calcium oscillations consistent with permanent photodynamic receptor activation.
Usefulness & Problems
Why this is useful
KillerRed is useful as a genetically encoded light-responsive source of singlet oxygen that can be targeted to defined subcellular locations. The cited work supports its use for optical activation of CCK1R signaling from the plasma membrane in living cells.
Source:
KillerRed, miniSOG, miniSOG2, singlet oxygen protein photosensitizer (SOPP), flavin-binding fluorescent protein from Methylobacterium radiotolerans with point mutation C71G (Mr4511C71G), and flavin-binding fluorescent protein from Dinoroseobacter shibae (DsFbFP) were expressed at the plasma membrane (PM) in AR4-2J cells, which express endogenous CCK1R. Light irradiation ... of GEPPPM-expressing AR4-2J was found to all trigger persistent calcium oscillations, a hallmark of permanent photodynamic CCK1R activation; DsFbFP was the least effective, due to poor expression.
Problem solved
KillerRed helps solve the problem of inducing receptor signaling with light through a genetically encoded photosensitizer rather than an exogenous chemical photosensitizer alone. In the cited context, it enabled photodynamic activation of CCK1R and downstream calcium oscillations in AR4-2J cells.
Source:
KillerRed, miniSOG, miniSOG2, singlet oxygen protein photosensitizer (SOPP), flavin-binding fluorescent protein from Methylobacterium radiotolerans with point mutation C71G (Mr4511C71G), and flavin-binding fluorescent protein from Dinoroseobacter shibae (DsFbFP) were expressed at the plasma membrane (PM) in AR4-2J cells, which express endogenous CCK1R. Light irradiation ... of GEPPPM-expressing AR4-2J was found to all trigger persistent calcium oscillations, a hallmark of permanent photodynamic CCK1R activation; DsFbFP was the least effective, due to poor expression.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level protein part used inside a larger architecture that realizes a mechanism.
Techniques
Selection / EnrichmentTarget processes
selectionInput: Light
Implementation Constraints
The evidence supports use of KillerRed as a genetically encoded construct with plasma membrane targeting for cellular photodynamic experiments. The tool was applied by light irradiation in AR4-2J cells, but the supplied evidence does not specify construct architecture, chromophore requirements, or illumination parameters.
The supplied evidence is limited to one study and one application context, namely photodynamic activation of CCK1R in AR4-2J cells. The provided claims do not report quantitative performance metrics for KillerRed, wavelength requirements, expression characteristics, or validation outside this receptor-signaling assay.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Source 2review2014Redox Biology
Source 3primary paper2020Biomolecules
Ranked Claims
CCK1R is activated by singlet oxygen generated in photodynamic action with SALPC or genetically encoded protein photosensitizers including KillerRed and miniSOG.
Cholecystokinin 1 receptor (CCK1R) is activated by singlet oxygen (1O2) generated in photodynamic action with sulphonated aluminum phthalocyanine (SALPC) or genetically encoded protein photosensitizer (GEPP) KillerRed or mini singlet oxygen generator (miniSOG).
CCK1R is activated by singlet oxygen generated in photodynamic action with SALPC or genetically encoded protein photosensitizers including KillerRed and miniSOG.
Cholecystokinin 1 receptor (CCK1R) is activated by singlet oxygen (1O2) generated in photodynamic action with sulphonated aluminum phthalocyanine (SALPC) or genetically encoded protein photosensitizer (GEPP) KillerRed or mini singlet oxygen generator (miniSOG).
CCK1R is activated by singlet oxygen generated in photodynamic action with SALPC or genetically encoded protein photosensitizers including KillerRed and miniSOG.
Cholecystokinin 1 receptor (CCK1R) is activated by singlet oxygen (1O2) generated in photodynamic action with sulphonated aluminum phthalocyanine (SALPC) or genetically encoded protein photosensitizer (GEPP) KillerRed or mini singlet oxygen generator (miniSOG).
CCK1R is activated by singlet oxygen generated in photodynamic action with SALPC or genetically encoded protein photosensitizers including KillerRed and miniSOG.
Cholecystokinin 1 receptor (CCK1R) is activated by singlet oxygen (1O2) generated in photodynamic action with sulphonated aluminum phthalocyanine (SALPC) or genetically encoded protein photosensitizer (GEPP) KillerRed or mini singlet oxygen generator (miniSOG).
CCK1R is activated by singlet oxygen generated in photodynamic action with SALPC or genetically encoded protein photosensitizers including KillerRed and miniSOG.
Cholecystokinin 1 receptor (CCK1R) is activated by singlet oxygen (1O2) generated in photodynamic action with sulphonated aluminum phthalocyanine (SALPC) or genetically encoded protein photosensitizer (GEPP) KillerRed or mini singlet oxygen generator (miniSOG).
CCK1R is activated by singlet oxygen generated in photodynamic action with SALPC or genetically encoded protein photosensitizers including KillerRed and miniSOG.
Cholecystokinin 1 receptor (CCK1R) is activated by singlet oxygen (1O2) generated in photodynamic action with sulphonated aluminum phthalocyanine (SALPC) or genetically encoded protein photosensitizer (GEPP) KillerRed or mini singlet oxygen generator (miniSOG).
CCK1R is activated by singlet oxygen generated in photodynamic action with SALPC or genetically encoded protein photosensitizers including KillerRed and miniSOG.
Cholecystokinin 1 receptor (CCK1R) is activated by singlet oxygen (1O2) generated in photodynamic action with sulphonated aluminum phthalocyanine (SALPC) or genetically encoded protein photosensitizer (GEPP) KillerRed or mini singlet oxygen generator (miniSOG).
KillerRed, miniSOG, miniSOG2, SOPP, Mr4511C71G, and DsFbFP expressed at the plasma membrane in AR4-2J cells all triggered persistent calcium oscillations upon light irradiation, consistent with permanent photodynamic CCK1R activation.
KillerRed, miniSOG, miniSOG2, singlet oxygen protein photosensitizer (SOPP), flavin-binding fluorescent protein from Methylobacterium radiotolerans with point mutation C71G (Mr4511C71G), and flavin-binding fluorescent protein from Dinoroseobacter shibae (DsFbFP) were expressed at the plasma membrane (PM) in AR4-2J cells, which express endogenous CCK1R. Light irradiation ... of GEPPPM-expressing AR4-2J was found to all trigger persistent calcium oscillations, a hallmark of permanent photodynamic CCK1R activation; DsFbFP was the least effective, due to poor expression.
KillerRed, miniSOG, miniSOG2, SOPP, Mr4511C71G, and DsFbFP expressed at the plasma membrane in AR4-2J cells all triggered persistent calcium oscillations upon light irradiation, consistent with permanent photodynamic CCK1R activation.
KillerRed, miniSOG, miniSOG2, singlet oxygen protein photosensitizer (SOPP), flavin-binding fluorescent protein from Methylobacterium radiotolerans with point mutation C71G (Mr4511C71G), and flavin-binding fluorescent protein from Dinoroseobacter shibae (DsFbFP) were expressed at the plasma membrane (PM) in AR4-2J cells, which express endogenous CCK1R. Light irradiation ... of GEPPPM-expressing AR4-2J was found to all trigger persistent calcium oscillations, a hallmark of permanent photodynamic CCK1R activation; DsFbFP was the least effective, due to poor expression.
KillerRed, miniSOG, miniSOG2, SOPP, Mr4511C71G, and DsFbFP expressed at the plasma membrane in AR4-2J cells all triggered persistent calcium oscillations upon light irradiation, consistent with permanent photodynamic CCK1R activation.
KillerRed, miniSOG, miniSOG2, singlet oxygen protein photosensitizer (SOPP), flavin-binding fluorescent protein from Methylobacterium radiotolerans with point mutation C71G (Mr4511C71G), and flavin-binding fluorescent protein from Dinoroseobacter shibae (DsFbFP) were expressed at the plasma membrane (PM) in AR4-2J cells, which express endogenous CCK1R. Light irradiation ... of GEPPPM-expressing AR4-2J was found to all trigger persistent calcium oscillations, a hallmark of permanent photodynamic CCK1R activation; DsFbFP was the least effective, due to poor expression.
KillerRed, miniSOG, miniSOG2, SOPP, Mr4511C71G, and DsFbFP expressed at the plasma membrane in AR4-2J cells all triggered persistent calcium oscillations upon light irradiation, consistent with permanent photodynamic CCK1R activation.
KillerRed, miniSOG, miniSOG2, singlet oxygen protein photosensitizer (SOPP), flavin-binding fluorescent protein from Methylobacterium radiotolerans with point mutation C71G (Mr4511C71G), and flavin-binding fluorescent protein from Dinoroseobacter shibae (DsFbFP) were expressed at the plasma membrane (PM) in AR4-2J cells, which express endogenous CCK1R. Light irradiation ... of GEPPPM-expressing AR4-2J was found to all trigger persistent calcium oscillations, a hallmark of permanent photodynamic CCK1R activation; DsFbFP was the least effective, due to poor expression.
KillerRed, miniSOG, miniSOG2, SOPP, Mr4511C71G, and DsFbFP expressed at the plasma membrane in AR4-2J cells all triggered persistent calcium oscillations upon light irradiation, consistent with permanent photodynamic CCK1R activation.
KillerRed, miniSOG, miniSOG2, singlet oxygen protein photosensitizer (SOPP), flavin-binding fluorescent protein from Methylobacterium radiotolerans with point mutation C71G (Mr4511C71G), and flavin-binding fluorescent protein from Dinoroseobacter shibae (DsFbFP) were expressed at the plasma membrane (PM) in AR4-2J cells, which express endogenous CCK1R. Light irradiation ... of GEPPPM-expressing AR4-2J was found to all trigger persistent calcium oscillations, a hallmark of permanent photodynamic CCK1R activation; DsFbFP was the least effective, due to poor expression.
KillerRed, miniSOG, miniSOG2, SOPP, Mr4511C71G, and DsFbFP expressed at the plasma membrane in AR4-2J cells all triggered persistent calcium oscillations upon light irradiation, consistent with permanent photodynamic CCK1R activation.
KillerRed, miniSOG, miniSOG2, singlet oxygen protein photosensitizer (SOPP), flavin-binding fluorescent protein from Methylobacterium radiotolerans with point mutation C71G (Mr4511C71G), and flavin-binding fluorescent protein from Dinoroseobacter shibae (DsFbFP) were expressed at the plasma membrane (PM) in AR4-2J cells, which express endogenous CCK1R. Light irradiation ... of GEPPPM-expressing AR4-2J was found to all trigger persistent calcium oscillations, a hallmark of permanent photodynamic CCK1R activation; DsFbFP was the least effective, due to poor expression.
KillerRed, miniSOG, miniSOG2, SOPP, Mr4511C71G, and DsFbFP expressed at the plasma membrane in AR4-2J cells all triggered persistent calcium oscillations upon light irradiation, consistent with permanent photodynamic CCK1R activation.
KillerRed, miniSOG, miniSOG2, singlet oxygen protein photosensitizer (SOPP), flavin-binding fluorescent protein from Methylobacterium radiotolerans with point mutation C71G (Mr4511C71G), and flavin-binding fluorescent protein from Dinoroseobacter shibae (DsFbFP) were expressed at the plasma membrane (PM) in AR4-2J cells, which express endogenous CCK1R. Light irradiation ... of GEPPPM-expressing AR4-2J was found to all trigger persistent calcium oscillations, a hallmark of permanent photodynamic CCK1R activation; DsFbFP was the least effective, due to poor expression.
DsFbFP was the least effective among the tested plasma-membrane-targeted genetically encoded photosensitizers because of poor expression.
DsFbFP was the least effective, due to poor expression.
DsFbFP was the least effective among the tested plasma-membrane-targeted genetically encoded photosensitizers because of poor expression.
DsFbFP was the least effective, due to poor expression.
DsFbFP was the least effective among the tested plasma-membrane-targeted genetically encoded photosensitizers because of poor expression.
DsFbFP was the least effective, due to poor expression.
DsFbFP was the least effective among the tested plasma-membrane-targeted genetically encoded photosensitizers because of poor expression.
DsFbFP was the least effective, due to poor expression.
DsFbFP was the least effective among the tested plasma-membrane-targeted genetically encoded photosensitizers because of poor expression.
DsFbFP was the least effective, due to poor expression.
DsFbFP was the least effective among the tested plasma-membrane-targeted genetically encoded photosensitizers because of poor expression.
DsFbFP was the least effective, due to poor expression.
DsFbFP was the least effective among the tested plasma-membrane-targeted genetically encoded photosensitizers because of poor expression.
DsFbFP was the least effective, due to poor expression.
Different plasma-membrane-targeted genetically encoded protein photosensitizers could all photodynamically activate CCK1R.
In conclusion, different GEPPPM could all photodynamically activate CCK1R.
Different plasma-membrane-targeted genetically encoded protein photosensitizers could all photodynamically activate CCK1R.
In conclusion, different GEPPPM could all photodynamically activate CCK1R.
Different plasma-membrane-targeted genetically encoded protein photosensitizers could all photodynamically activate CCK1R.
In conclusion, different GEPPPM could all photodynamically activate CCK1R.
Different plasma-membrane-targeted genetically encoded protein photosensitizers could all photodynamically activate CCK1R.
In conclusion, different GEPPPM could all photodynamically activate CCK1R.
Different plasma-membrane-targeted genetically encoded protein photosensitizers could all photodynamically activate CCK1R.
In conclusion, different GEPPPM could all photodynamically activate CCK1R.
Different plasma-membrane-targeted genetically encoded protein photosensitizers could all photodynamically activate CCK1R.
In conclusion, different GEPPPM could all photodynamically activate CCK1R.
Different plasma-membrane-targeted genetically encoded protein photosensitizers could all photodynamically activate CCK1R.
In conclusion, different GEPPPM could all photodynamically activate CCK1R.
Devazepide at 2 nM readily inhibited LED-induced calcium oscillations in miniSOG plasma-membrane-targeted AR4-2J cells, was less effective in mitochondria-targeted miniSOG cells, and did not inhibit lysosome-targeted miniSOG cells.
In miniSOGPM-AR4-2J cells, light emitting diode (LED) light irradiation-induced calcium oscillations were readily inhibited by CCK1R antagonist devazepide 2 nM; miniSOGMT-AR4-2J cells were less susceptible, but miniSOGLS-AR4-2J cells were not inhibited.
devazepide concentration 2 nM
Devazepide at 2 nM readily inhibited LED-induced calcium oscillations in miniSOG plasma-membrane-targeted AR4-2J cells, was less effective in mitochondria-targeted miniSOG cells, and did not inhibit lysosome-targeted miniSOG cells.
In miniSOGPM-AR4-2J cells, light emitting diode (LED) light irradiation-induced calcium oscillations were readily inhibited by CCK1R antagonist devazepide 2 nM; miniSOGMT-AR4-2J cells were less susceptible, but miniSOGLS-AR4-2J cells were not inhibited.
devazepide concentration 2 nM
Devazepide at 2 nM readily inhibited LED-induced calcium oscillations in miniSOG plasma-membrane-targeted AR4-2J cells, was less effective in mitochondria-targeted miniSOG cells, and did not inhibit lysosome-targeted miniSOG cells.
In miniSOGPM-AR4-2J cells, light emitting diode (LED) light irradiation-induced calcium oscillations were readily inhibited by CCK1R antagonist devazepide 2 nM; miniSOGMT-AR4-2J cells were less susceptible, but miniSOGLS-AR4-2J cells were not inhibited.
devazepide concentration 2 nM
Devazepide at 2 nM readily inhibited LED-induced calcium oscillations in miniSOG plasma-membrane-targeted AR4-2J cells, was less effective in mitochondria-targeted miniSOG cells, and did not inhibit lysosome-targeted miniSOG cells.
In miniSOGPM-AR4-2J cells, light emitting diode (LED) light irradiation-induced calcium oscillations were readily inhibited by CCK1R antagonist devazepide 2 nM; miniSOGMT-AR4-2J cells were less susceptible, but miniSOGLS-AR4-2J cells were not inhibited.
devazepide concentration 2 nM
Devazepide at 2 nM readily inhibited LED-induced calcium oscillations in miniSOG plasma-membrane-targeted AR4-2J cells, was less effective in mitochondria-targeted miniSOG cells, and did not inhibit lysosome-targeted miniSOG cells.
In miniSOGPM-AR4-2J cells, light emitting diode (LED) light irradiation-induced calcium oscillations were readily inhibited by CCK1R antagonist devazepide 2 nM; miniSOGMT-AR4-2J cells were less susceptible, but miniSOGLS-AR4-2J cells were not inhibited.
devazepide concentration 2 nM
Devazepide at 2 nM readily inhibited LED-induced calcium oscillations in miniSOG plasma-membrane-targeted AR4-2J cells, was less effective in mitochondria-targeted miniSOG cells, and did not inhibit lysosome-targeted miniSOG cells.
In miniSOGPM-AR4-2J cells, light emitting diode (LED) light irradiation-induced calcium oscillations were readily inhibited by CCK1R antagonist devazepide 2 nM; miniSOGMT-AR4-2J cells were less susceptible, but miniSOGLS-AR4-2J cells were not inhibited.
devazepide concentration 2 nM
Devazepide at 2 nM readily inhibited LED-induced calcium oscillations in miniSOG plasma-membrane-targeted AR4-2J cells, was less effective in mitochondria-targeted miniSOG cells, and did not inhibit lysosome-targeted miniSOG cells.
In miniSOGPM-AR4-2J cells, light emitting diode (LED) light irradiation-induced calcium oscillations were readily inhibited by CCK1R antagonist devazepide 2 nM; miniSOGMT-AR4-2J cells were less susceptible, but miniSOGLS-AR4-2J cells were not inhibited.
devazepide concentration 2 nM
miniSOG targeted to the plasma membrane, mitochondria, or lysosomes in AR4-2J cells induced persistent calcium oscillations after LED light irradiation.
miniSOG was targeted to PM, mitochondria (MT) or lysosomes (LS) in AR4-2J in parallel experiments; LED light irradiation was found to all induce persistent calcium oscillations.
miniSOG targeted to the plasma membrane, mitochondria, or lysosomes in AR4-2J cells induced persistent calcium oscillations after LED light irradiation.
miniSOG was targeted to PM, mitochondria (MT) or lysosomes (LS) in AR4-2J in parallel experiments; LED light irradiation was found to all induce persistent calcium oscillations.
miniSOG targeted to the plasma membrane, mitochondria, or lysosomes in AR4-2J cells induced persistent calcium oscillations after LED light irradiation.
miniSOG was targeted to PM, mitochondria (MT) or lysosomes (LS) in AR4-2J in parallel experiments; LED light irradiation was found to all induce persistent calcium oscillations.
miniSOG targeted to the plasma membrane, mitochondria, or lysosomes in AR4-2J cells induced persistent calcium oscillations after LED light irradiation.
miniSOG was targeted to PM, mitochondria (MT) or lysosomes (LS) in AR4-2J in parallel experiments; LED light irradiation was found to all induce persistent calcium oscillations.
miniSOG targeted to the plasma membrane, mitochondria, or lysosomes in AR4-2J cells induced persistent calcium oscillations after LED light irradiation.
miniSOG was targeted to PM, mitochondria (MT) or lysosomes (LS) in AR4-2J in parallel experiments; LED light irradiation was found to all induce persistent calcium oscillations.
miniSOG targeted to the plasma membrane, mitochondria, or lysosomes in AR4-2J cells induced persistent calcium oscillations after LED light irradiation.
miniSOG was targeted to PM, mitochondria (MT) or lysosomes (LS) in AR4-2J in parallel experiments; LED light irradiation was found to all induce persistent calcium oscillations.
miniSOG targeted to the plasma membrane, mitochondria, or lysosomes in AR4-2J cells induced persistent calcium oscillations after LED light irradiation.
miniSOG was targeted to PM, mitochondria (MT) or lysosomes (LS) in AR4-2J in parallel experiments; LED light irradiation was found to all induce persistent calcium oscillations.
miniSOG is associated with correlative light and electron microscopy applications in the supplied source scaffold.
SuperNova and miniSOG are associated with chromophore-assisted light inactivation workflows in the supplied source scaffold.
At the organelle level, including mitochondria, plasma membrane, or lysosomes, CALI can trigger cell death.
CALI can provide information about individual events involved in target protein function and highlight them within multifactorial events.
CALI has emerged as an optogenetic tool to switch off signaling pathways, including optical depletion of individual neurons.
CALI of nuclear proteins can induce cell cycle arrest and chromatin- or locus-specific DNA damage.
Rescue experiments can clarify phenotypic capabilities after CALI depletion of endogenous targets.
Using spatially restricted microscopy illumination, CALI can address protein isoform, subcellular localization, and phase-specific questions that RNA interference or chemical treatment cannot.
CALI is performed using photosensitizers that generate reactive oxygen species.
CALI enables spatiotemporal knockdown or loss-of-function of target molecules in situ.
The review describes two CALI approaches: transgenic tags with chemical photosensitizers and genetically encoded fluorescent protein fusions.
This review centers on genetically encoded ROS-generating proteins for optogenetic control of reactive oxygen species, with KillerRed, miniSOG, and SuperNova highlighted as core examples.
Approval Evidence
3 sources4 linked approval claimsfirst-pass slug killerred
genetically encoded protein photosensitizer (GEPP) KillerRed
Source:
Additional high-signal enrichment leads cluster into four useful categories: foundational CALI methodology, mechanistic papers explaining ROS-mediated inactivation, genetically encoded photosensitizer/tool-development papers (notably KillerRed, miniSOG, SuperNova), and representative application papers in neurons, mitochondria, nuclei, and whole-animal cell ablation.
Source:
The anchor review explicitly centers on genetically encoded ROS-generating proteins used for optogenetic control of reactive oxygen species, especially KillerRed, miniSOG, and the then-new monomeric derivative SuperNova.
Source:
activationsupports
CCK1R is activated by singlet oxygen generated in photodynamic action with SALPC or genetically encoded protein photosensitizers including KillerRed and miniSOG.
Cholecystokinin 1 receptor (CCK1R) is activated by singlet oxygen (1O2) generated in photodynamic action with sulphonated aluminum phthalocyanine (SALPC) or genetically encoded protein photosensitizer (GEPP) KillerRed or mini singlet oxygen generator (miniSOG).
Source:
application resultsupports
KillerRed, miniSOG, miniSOG2, SOPP, Mr4511C71G, and DsFbFP expressed at the plasma membrane in AR4-2J cells all triggered persistent calcium oscillations upon light irradiation, consistent with permanent photodynamic CCK1R activation.
KillerRed, miniSOG, miniSOG2, singlet oxygen protein photosensitizer (SOPP), flavin-binding fluorescent protein from Methylobacterium radiotolerans with point mutation C71G (Mr4511C71G), and flavin-binding fluorescent protein from Dinoroseobacter shibae (DsFbFP) were expressed at the plasma membrane (PM) in AR4-2J cells, which express endogenous CCK1R. Light irradiation ... of GEPPPM-expressing AR4-2J was found to all trigger persistent calcium oscillations, a hallmark of permanent photodynamic CCK1R activation; DsFbFP was the least effective, due to poor expression.
Source:
conclusionsupports
Different plasma-membrane-targeted genetically encoded protein photosensitizers could all photodynamically activate CCK1R.
In conclusion, different GEPPPM could all photodynamically activate CCK1R.
Source:
review scope summarysupports
This review centers on genetically encoded ROS-generating proteins for optogenetic control of reactive oxygen species, with KillerRed, miniSOG, and SuperNova highlighted as core examples.
Source:
Comparisons
Source-backed strengths
The source literature identifies KillerRed as one of several genetically encoded photosensitizers that activated CCK1R through singlet oxygen generation. When expressed at the plasma membrane in AR4-2J cells, it triggered persistent calcium oscillations upon irradiation, indicating effective photodynamic signaling activation.
Source:
DsFbFP was the least effective, due to poor expression.
Ranked Citations
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