Biosensor expression plasmids
Our previously published plasmids are available from Addgene, a non-profit plasmid repository:
Plasmids for bacterial expression
roGFP2-His in pQE-60
https://www.addgene.org/65046
Grx1-roGFP2-His in pQE-60 [1]
https://www.addgene.org/64799
roGFP2-Orp1-His in pQE-60 [2]
https://www.addgene.org/64976
Plasmids for yeast expression
cyto-roGFP2-yGrx1 in p415TEF [4]
https://www.addgene.org/65004
cyto-roGFP2-yGrx2 in p415TEF [4]
https://www.addgene.org/65005
cyto-roGFP1-iL-hGrx1 in p415TEF [4]
https://www.addgene.org/65002
cyto-roGFP2-Tsa2ΔCR in p415TEF [6]
https://www.addgene.org/83238
cyto-roGFP2-Tsa2ΔCPΔCR in p415TEF [6]
https://www.addgene.org/83239
Plasmids for insect expression
cyto-Grx1-roGFP2 in pUAST [3]
https://www.addgene.org/64994
mito-roGFP2-Grx1 in pUAST [3][5]
https://www.addgene.org/64995
cyto-roGFP2-Orp1 in pUAST [3]
https://www.addgene.org/64996
mito-roGFP2-Orp1 in pUAST 3]
https://www.addgene.org/64997
cyto-Grx1-roGFP2 in pCaSpeR4:alphaTub84B[3]
https://www.addgene.org/64999
mito-roGFP2-Grx1 in pCaSpeR4:alphaTub84B [3][5]
https://www.addgene.org/65000
cyto-roGFP2-Orp1 in pCaSpeR4:alphaTub84B [3]
https://www.addgene.org/64998
mito-roGFP2-Orp1 in pCaSpeR4:alphaTub84B [5]
https://www.addgene.org/65001
Plasmids for mammalian expression
cyto-Grx1-roGFP2 in pLPCX (retroviral) [1]
https://www.addgene.org/64975
cyto-roGFP2-Orp1 in pLPCX (retroviral) [2]
https://www.addgene.org/64991
mito-Grx1-roGFP2 in pLPCX (retroviral) [1]
https://www.addgene.org/64977
mito-roGFP2-Orp1 in pLPCX (retroviral) [2]
https://www.addgene.org/64992
cyto-Grx1-roGFP2 in pEIGW (lentiviral) [1]
https://www.addgene.org/64993
Original probe references
[1] Gutscher et al. (2008) Real-time imaging of the intracellular glutathione redox potential. Nat Methods 5: 553-9
[2] Gutscher et al. (2009) Proximity-based protein thiol oxidation by H2O2-scavenging peroxidases. J Biol Chem 284: 31532-40
[3] Albrecht et al. (2011) In vivo mapping of hydrogen peroxide and oxidized glutathione reveals chemical and regional specificity of redox homeostasis. Cell Metab 14: 819-829
[4] Morgan et al. (2013) Multiple glutathione disulfide removal pathways mediate cytosolic redox homeostasis. Nat Chem Biol 9:119-125
[5] Albrecht et al. (2014) Redesign of genetically-encoded biosensors for monitoring mitochondrial redox status in a broad range of model eukaryotes. J Biomol Screen 19:379-86
[6] Morgan et al. (2016) Real-time monitoring of basal H2O2 levels with peroxiredoxin-based probes. Nature Chemical Biology 12: 437-443
Probe reviews
For detailed background information about roGFP2-based redox probes (mode of operation, limitations etc.):
Meyer and Dick (2010) Fluorescent Protein-based Redox Probes. Antioxid Redox Signal 13: 621-650
Ezeriņa et al. (2014) Imaging dynamic redox processes with genetically encoded probes. J Mol Cell Cardiol 73:43-49
Schwarzländer et al. (2015) Dissecting Redox Biology using Fluorescent Protein Sensors. Antioxid Redox Signal 24:680-712
For practical information on how to perform measurements (microscopy settings etc.) and data analysis:
Morgan et al. (2011) Measuring EGSH and H2O2 with roGFP2-based redox probes. Free Radic Biol Med 51: 1943-1951
Barata and Dick (2013) In Vivo Imaging of H2O2 Production in Drosophila. Meth Enzymol 526:61-82