Fluorescent Proteins: Biosensors

Background

Genetically encoded fluorescent biosensors are fluorescent proteins attached to an additional protein sequence that make it sensitive to small biomolecules or other physiological intracellular processes. Fluorescent biosensors are introduced into cells, tissues or organisms to allow for detection by fluorescence microscopy as a difference in FRET efficiency, translocation of the fluorescent protein or modulation of the fluorescent properties of a single fluorescent protein. Many biosensors allow for long-term imaging and can be designed to specifically target cellular compartments or organelles. Another advantage of biosensors is that they permit investigation of a signaling pathway or measurement of a biomolecule while largely preserving spatial and temporal cellular processes.

The following plasmids encode fluorescent biosensors that were designed to measure specific small biomolecules or the activity of specific genes via circularly permuted fluorescent protein (cpFP) and/or FRET-based fluorescent protein constructs. Custom biosensors can also be constructed using the cpFRET kit from the Pertz laboratory.

Small Molecule Biosensors

Category	Purpose	Principal Investigator	Plasmids
Acetylcholine	Family of ACh sensors (GACh) based on G-protein-coupled receptors	Yulong Li	See Plasmids
Arabinose	Monitor intracellular levels of arabinose	Wolf Frommer	See Plasmid
ATP:ADP	Monitor the ATP:ADP ratio during live-cell imaging	Gary Yellen	See Plasmids
ATP	Multicolor ATP indicators	Tetsuya Kitaguchi	See Plasmids
Calcium	Large palette of genetically encoded Ca2+ indicators (GECOs)	Robert Campbell	See Plasmids
Calcium	tdTomato-mAmetrine1.2 Ca2+ sensors	Robert Campbell	See Plasmids
Calcium	Calcium indicator based on C-terminal domain of Opsanus troponin C	Oliver Griesbeck	See Plasmids
Calcium	High-performing calcium indicator based on C-terminal domain of troponin C	Oliver Griesbeck	See Plasmids
Calcium	Calcium indicator for neural activity imaging (GCaMP5G)	Loren Looger & Douglas Kim	See Plasmids
Calcium	Updated calcium sensors for imaging neural activity (GCaMP6)	Douglas Kim	See Plasmids
Calcium	Fast response calcium sensors (GCaMP3-derived)	Katalin Torok	See Plasmids
cAMP (cyclic AMP)	Improved yellow fluorescent cAMP indicator (Flamindo2)	Tetsuya Kitaguchi	See Plasmids
cAMP (cyclic AMP)	Red fluorescent protein-based cAMP indicator (Pink Flamindo)	Tetsuya Kitaguchi	See Plasmid
cGMP (cyclic GMP)	Green fluorescent probe for cGMP	Tetsuya Kitaguchi	See Plasmid
Dopamine	Genetically encoded GPCR-activation-based-DA (GRABDA) sensors	Yulong Li	See Plasmids
Glucose	Monitor glucose levels and transport	Wolf Frommer	See Plasmids
Glucose	Green fluorescent glucose indicators for living cells (Glifons)	Tetsuya Kitaguchi	See Plasmids
Glutamate	Visualize glutamate neurotransmission	Loren Looger	See Plasmids
Glutamate	Monitor glutamate transport and metabolism	Wolf Frommer	See Plasmids
Glutamate	Ultrafast glutamate sensors	Katalin Torok	See Plasmids
Hydrogen Peroxide (H2O2)	Measure changes in intracellular hydrogen peroxide concentration	Vsevolod Belousov	See Plasmids
Lactate	Measure lactate concentration and lactate fluxes	Luis Felipe Barros	See Plasmids
Magnesium	Fluorescent Mg2+ sensor	Maarten Merkx	See Plasmids
Maltose	Monitor maltose levels and metabolism	Wolf Frommer	For Bacteria For Plants
NADH-NAD+	Fluorescent biosensor of the cytosolic NADH-NAD+ redox state	Vsevolod Belousov	See Plasmids
NADH-NAD+	Fluorescent biosensor of the cytosolic NADH-NAD+ redox state	Gary Yellen	See Plasmids
Norepinephrine	Family of GPCR activation-based NE (GRABNE) sensors	Yulong Li	See Plasmids
pH	Image intracellular pH in live cells	Gary Yellen	See Plasmids
pH	pH biosensor to detect concentrative nucleoside transport	Robert Campbell	See Plasmids
pH	Fluorescent pH biosensor	Vsevolod Belousov	See Plasmids
pH	Mitochondrial pH biosensor	Nicolas Demaurex	See Plasmids
Phosphate	Monitor inorganic phosphate transport and metabolism	Wolf Frommer	See Plasmids
PIP3 & H2O2	Dual PIP3 and hydrogen peroxide biosensor	Vsevolod Belousov	See Plasmid
redox/ROS	Redox-sensitive cytosolic protein sensor	Ulrich Hammerling	See Plasmids
Ribose	Monitor ribose uptake and metabolism	Wolf Frommer	See Plasmids
Sucrose	Monitor sucrose levels and flux	Wolf Frommer	See Plasmids
Tryptophan	Visualize intracellular tryptophan levels and flux	Wolf Frommer	See Plasmids
Voltage	Fast fluorescent protein voltage sensor	Thomas Knopfel	See Plasmids
Voltage	Detection of single action / excitatory potentials in individual neurons and dendrites	Vincent Pieribone	See Plasmids
Voltage	Ultrafast fluorescent voltage sensor for detection of single action potentials	Michael Lin	See Plasmids
Zinc	Visualize free zinc levels in live cells	Maarten Merkx	See Plasmids
Zinc	Robust red zinc sensors	Maarten Merkx	See Plasmids

Gene-specific Biosensors

Gene	Purpose	Principal Investigator	Plasmids
Actin	Quantify actin polymerization with mEGFP-actin and sREACh-actin monomers	Ryohei Yasuda	See Plasmids
Arf6	Visualize Arf6 activation in live cells	Martin Schwartz	See Plasmids
Aurora B	Sensor for Aurora B substrate phosphorylation	Daniel Gerlich	See Plasmids
CaMKIIa	Measure CaMKII activation in dendritic spines	Ryohei Yasuda	See Plasmids
Caspase 3, 8/10	Reporter proteins for caspase activity	Peter Sorger	See Plasmids
Caspase 3	Reporter protein for caspase 3 activity	Robert Campbell	See Plasmid
Caspase 3	Reporter protein for caspase 3 activity	Vladislav Verkhusha	See Plasmid
Cdt	NIR reporter for cell cycle progression	Vladislav Verkhusha	See Plasmid
CyclinB-Cdk1	CyclinB1-Cdk1 kinase activity	Jonathon Pines	See Plasmids
GAT1	Detect assembly and oligomerization of GABA transporter (GAT1)	Henry Lester	See Plasmid
Geminin	NIR reporter for cell cycle progression	Vladislav Verkhusha	See Plasmid
IκBα	NIR reporter for canonical activation of NF-κB	Vladislav Verkhusha	See Plasmid
Ras GTPase	Sensors for the activation of Ras GTPase with slow (FRas) and fast (FRas-F) kinetics	Karel Svoboda	See Plasmids
Rho GTPases	Biosensors for characterizing dynamics of Rho family GTPases in living cells	Klaus Hahn	See Plasmids
RhoA	Visualize RhoA activity during cell migration	Klaus Hahn	See Plasmids