PFD-rAAV-TRE3g-jGCaMP7s-WPRE-hGH polyA

Promoter Details

TRE3G, tetracycline-induced expression, 350bp.

Functional Gene Details

jGCaM P7: By optimizing the Ca2+ affinity of a calcium ion probe while further improving the probe's sensitivity, signal-to-noise ratio, half-life, and dynamic range, Karel Svoboda, Loren Looger, and Vivek from the Howard Hughes Medical Institute Jayaraman and other research groups jointly developed the jGCaMP7 series, including: jGCaMP7s (sensitive), jGCaMP7f (fast kinetics), jGCaMP7b (brighter baseline fluorescence) and jGCaMP7c (high contrast), which are specially used to detect neuronal activity in specific environments. Tracking the dynamics of neuronal subcells is of great significance to the study of neural circuits. However, subcellular structures such as dendrites and axons are small, and jGCaMP6 has low sensitivity and low imaging fidelity. However, jGCaMP7s, jGCaMP7f, and jGCaMP7b have greatly improved the sensitivity under different calcium ion concentrations. jGCaMP7f also has a faster response rate, which is very important for the dynamic detection of neuronal subcells. jGCaMP7 was tested both in vivo and in vitro and performed significantly better than GCaMP6. The sensitivity of jGCaMP7s is more than 5 times that of GCaMP6s, and the speed is faster than GCaMP6s; the sensitivity of jGCaMP7b is 3 times that of GCaMP6s, and the fluorescence brightness is enhanced by 50%; the sensitivity of jGCaMP7c is 2.7 times that of GCaMP6s, and the contrast is enhanced; the reaction rate of jGCaMP7f is 5 times that of GCaMP6f and 3 times that of GCaMP6s. jGCaMP7s more sensitive jGCaMP7f fast kinetics jGCaMP7b high background jGCaMP7c high contrast, low background

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.