PFD-rAAV-CaMKIIa-mito-NES-jRGECO1a-WPRE-hGH polyA

Promoter Details

CaMKIIa, pyramidal neuron/excitatory neuron promoter, 1293bp.

Functional Gene Details

jRGECO1a: Red-shifted calcium imaging, fluorescence from mApple, NES-RCaMP1h variant 1519, active in mice, flies, fish and worms, using deep brain imaging, can be used simultaneously with the GCaMP green fluorescence series to study the activity and interactions of different types of neurons. The disadvantage is that existing red fluorescent detectors are not as useful as green ones. It can be used to study deep tissues by reducing scattering and tissue absorption, thereby reducing phototoxicity. The excitation wavelength is 1040nm or 1100nm. NES: export nuclear signals, nuclear export signal sequence, only distributed outside the nucleus and located in the cytoplasm. mito: mitochondrial localization.

jRGECO1a: Red-shifted calcium imaging, fluorescence from mApple, NES-RCaMP1h variant 1519, active in mice, flies, fish and worms, using deep brain imaging, can be used simultaneously with the GCaMP green fluorescence series to study the activity and interactions of different types of neurons. The disadvantage is that existing red fluorescent detectors are not as useful as green ones. It can be used to study deep tissues by reducing scattering and tissue absorption, thereby reducing phototoxicity. The excitation wavelength is 1040nm or 1100nm. NES: export nuclear signals, nuclear export signal sequence, only distributed outside the nucleus and located in the cytoplasm.

Application

Measuring neuronal activity using genetically encoded calcium indicators (GECIs) has become a widely used method in neuroscience. Calcium imaging with GECIs is often used to measure neural activity in the complete nervous system, such as detecting the activity of a large number of neuronal cell bodies, or tracking the dynamics of neuronal subcells (such as axons, dendrites, and single synapses). Calcium imaging system is a type of optogenetic technology that uses calcium ion indicators that can sense Ca2+ concentration to detect Ca2+ concentration changes in cells or tissues and convert calcium concentration changes into fluorescent signals, thereby converting cell electrical activity into recordable light signals.

Notes

1. Short-term storage at 4°C; long-term storage at -80°C

2. It can be stored at -80°C for more than 6 months. Each time it is frozen and thawed, the titer will be reduced by 10%, and repeated freezing and thawing should be avoided; if the virus is stored for more than 6 months, it is recommended to re-measure the virus titer before use

3. According to the experimental needs, the virus can be appropriately diluted and divided into the volume required for each experiment.