Cat.No. : AAB0064
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV Serotype 9 Storage: -80 ℃
| Cat. No. | AAB0064 |
| Description | Premade AAV particles in serotype 9 containing GCaMP6f under the control of a Syn promoter. |
| Serotype | AAV Serotype 9 |
| Tag | GCaMP6f |
| Product Type | Adeno-associated virus particles |
| Biosensor | GCaMP6f-Improved SNR, faster kinetics; Green indicator |
| Titer | Varies lot by lot, typically ≥1x10^12 GC/mL |
| Size | Varies lot by lot, for example, 30 μL, 50 μL, 100 μL etc. |
| Storage | Store at -80℃. Avoid multiple freeze/thaw cycles. |
| Shipping | Frozen on dry ice |
| Creative Biogene ensures high-quality AAV particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between AAV particle lots. | |
| Endotoxin | Endotoxins, primarily derived from Gram-negative bacteria, can trigger adverse immune responses. Endotoxin contamination is a significant concern in the production of AAV, especially for applications in animal studies and gene therapy. Effective endotoxin quality control is essential in the development and manufacturing of AAV particles. Creative Biogene utilizes rigorous endotoxin detection methods to monitor the endotoxin level in our produced AAV particles to ensure regulatory compliance. |
| Purity | AAV purity is critical for ensuring the safety and efficacy of AAV-based applications.AAV capsids are composed of three main protein components, known as viral proteins: VP1, VP2, and VP3. These proteins play a critical role in the structure and functionality of the AAV capsid. Monitoring the VP1, VP2, and VP3 content in AAV preparations is essential for quality control in AAV production. Our AAV particles are tested for showing three clear bands of VP1, VP2 VP3 by SDS-PAGE. |
| Sterility | The AAV virus samples are inoculated into the cell culture medium for about 5 days to detect bacterial and fungal growth. |
| Transducibility | Upon requirement, Creative Biogene can perform in vitro or in vivo transduction assays to evaluate the ability of AAV to deliver genetic material into target cells or tissues, and assess gene expression and functional activities. |
| Empty vs. Full Capsids | Based-on our proprietary AAV production and purification technology, Creative Biogene can always offer AAV particles with high ratio of full capsids. If required, we can also assess the ratio for a specifc lot of AAV particles by transmission electron microscopy (TEM) or other methods. |
Neural stem/progenitor cell grafts integrate into the site of spinal cord injury (SCI) and form anatomical and electrophysiological neuronal relays at the lesion. To determine how the grafts organize synaptically and connect with the host system, calcium imaging of neural progenitor cell grafts within the SCI site was performed using in vivo imaging and spinal cord slices. The stem cell grafts organized into spontaneously active local synaptic networks. Following optogenetic stimulation of the corticospinal tract axons that regenerated into the graft host, distinct and separate neuronal networks throughout the graft responded. Furthermore, optogenetic stimulation of graft axons extending from the lesion into the denervated spinal cord also triggered responses in local host neuronal networks. In vivo imaging revealed that behavioral stimulation of the host elicited focal synaptic responses within the graft. Thus, it is noteworthy that the neural progenitor cell grafts formed functional synaptic subnetworks in a pattern parallel to the normal spinal cord.
The researchers mixed the AAV-Syn-FLEX-ChrimsonR-tdTomato vector with Syn1-Cre neural progenitor cells, which were then acutely transplanted into T12 dorsal column lesions. This resulted in graft-specific expression of ChrimsonR-tdTomato, which was readily detected in axons extending from the graft border to the distal host spinal cord (Figures 1A-1B). Three to four weeks after transplantation, AAV9-Syn-GCaMP6f was injected into the host central gray matter 1.2 mm caudal to the graft (Figures 1A-1B). When imaged 6-8 weeks after transplantation, a total of 15 host neurons were detected in sections from two animals that responded to full-field stimulation of graft axons (Figures 1C-1F). Responding cells were located in the medial and dorsal gray matter. Host responses to graft axon stimulation were abolished after DNQX administration, indicating that host responses to graft axon stimulation were mediated through excitatory synaptic transmission (Figure 1G). The latency of these responses was less than 200 ms, indicating that, as with the grafts, the responses peaked before the end of the 500 ms light stimulation (Figure 1G). Thus, the neural progenitor cell grafts were able to activate host neurons beneath the spinal cord lesion.
Figure 1. Host neurons are activated by optogenetic stimulation of axons extending from grafts. (Ceto S, et al., 2019)
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