PFD-rAAV-EF1a-fDIO-GCaMP6m-Xc-WPRE-hGH polyA

Promoter Details

Ef1α, a strong promoter, human targeted elongation factor 1α (EF1A) gene, can be stably expressed in most types of cells, especially cells where the CMV promoter is silenced, such as stem cells, 1178bp.

Functional Gene Details

GCaMP-X: Research by Liu Xiaodong's research group found that apoCaM (non-calcium-bound calmodulin) severely interferes with the excitation-transcription coupling mediated by neuronal L-type voltage-gated calcium channels, causing abnormalities in neuronal calcium dynamics and gene expression. A specific apoCaM protective element was introduced at the nitrogen terminus of the GCaMP probe to synthesize a calcium ion probe, which can overcome the interference of apoCaM while still showing the excellent Ca2+ sensing properties of GCaMP. GCaMP-X has more advantages in long-term calcium ion monitoring. The non-destructive probe GCaMP-X effectively solves the shortcomings of traditional GCaMP probes and is capable of safe, non-destructive, long-term, accurate, specific and quantitative calcium signal monitoring of multiple organelles. This research also has guiding significance for the design of future CaM-based molecular tools. GCaMP-X requires a high titer (10^13vg/mL), and its characteristics include: 1. It does not accumulate in the nucleus; 2. It does not affect calmodulin-related pathways and does not damage neurons; 3. It can observe calcium signals for an extremely long time (≥4 weeks) (Our products are exclusively authorized by Liu Xiaodong’s research group at Beijing University of Aeronautics and Astronautics! Welcome to consult!). GCaMP6m-Xc represents cytoplasmic localization.

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.

Regulatory Element Details

fDIO: It is an element that cooperates with Flp to control the expression of target genes. If there is no Flp recombinase, the target gene will not be expressed. Only in the presence of Flp can the target gene be expressed normally.

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.