rsCaMPARI

The rsCaMPARI technology is a groundbreaking sensor technology that provides a reversible method for marking and tracking neuronal activity. Based on photoconvertible fluorescent proteins, traditional neuronal activity markers offer permanent and irreversible marking, which limits their utility in experiments requiring multiple snapshots of activity within the same sample. To address this limitation, rsCaMPARI employs a reversibly switchable fluorescent protein (rsFP) that can photoswitch between bright and dim states in response to different wavelengths of light. This innovative approach allows researchers to repeatedly erase and remark neuronal activity, making it ideal for dynamic and comparative studies.

The key to rsCaMPARI's functionality is the insertion of calcium-binding domains into the coding sequence of a reversibly switchable mutant of the fluorescent protein EosFP. This modification enables the photoswitching kinetics to be modulated by neuronal activity, with calcium acting as a crucial mediator. When neuronal activity causes a rise in calcium levels, rsCaMPARI undergoes rapid photoswitching, marking the active neurons. This marker can then be reset by light, allowing for repeated rounds of marking and observation. In practical applications, rsCaMPARi was expressed successfully in cultured rat hippocampal neurons, demonstrating reliable and reversible activity marking.

rsCaMPARI provides a versatile and efficient tool for neuroscientists to study neuronal activity with high temporal resolution. Its ability to reset and reuse the marker offers significant advantages for experiments requiring ongoing neural dynamics monitoring under varying conditions. By enabling repeated and precise tracking of neuronal activity, rsCaMPARI holds great potential for advancing our understanding of brain function and neural circuitry.
 

rsCaMPARI features: 
• Enables spatiotemporally precise marking, erasing, and remarking of active neuron populations 
• Works under under brief, user-defined time windows of light exposure 
• Kinetics are modulated by calcium concentration with blue light 
• Reset the fluorescence with violet light 
• Proof of action in freely swimming zebrafish 

Available Plasmids: 
• pRSET_His-rsCaMPARI-mRuby3 
• pAAV-hsyn_NES-His-rsCaMPARI-mRuby3 
• pAAV-hsyn_NLS-His-rsCaMPARI-mRuby3 
• pTol2-elavl3_NES-rsCaMPARI-mRuby3 

Opportunity: 

Available for commercial licensing (internal research or commercial sale). Academic researchers may access the existing materials via Addgene.

Intellectual Property:

Allowed US patent application: 17/040,856