Fluorescent Protein Guide: 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 |
Do you have suggestions for other plasmids that should be added to this list?
Fill out our Suggest a Plasmid form or e-mail [email protected] to help us improve this resource!