Cat.No. : AAV00191Z
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV Serotype 5 Storage: -80 ℃
| Cat. No. | AAV00191Z |
| Description | AAV serotype 5 particles contain calcium indicator GCaMP6s under CAG promoter. |
| Serotype | AAV Serotype 5 |
| 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. |
The trigeminal sensory innervation of the cranial meninges is thought to have nociceptive functions and mediate headaches. However, the activity of meningeal afferents under natural conditions in awake animals remains unexplored. Here, researchers used two-dimensional and three-dimensional two-photon calcium imaging to track the activity of meningeal afferent fibers in awake mice. Surprisingly, a large number of afferents were activated under non-noxious conditions such as locomotion. They estimated locomotion-associated meningeal deformations and found afferents with different dynamics and adapted to different degrees of meningeal expansion, compression, shear, and Z-axis motion. Moreover, these mechanosensitive afferents often adapted to different directions of meningeal expansion or compression. Thus, in addition to their role in headache-related pain, meningeal sensory neurons track the dynamic mechanical state of the meninges under natural conditions.
Here, researchers developed a calcium imaging method to simultaneously monitor the activity of dozens of meningeal afferent peripheral nerve terminals within the intact cranial cavity of awake, behaving mice through chronically implanted cranial windows. Microinjection of AAV5-CAG-GCaMP6 into the TG resulted in labeling of TG neuronal somata and afferent fibers in the ipsilateral dural meninges (Figures 1A-1C). GCaMP6 colocalized with calcitonin gene-related peptide (CGRP) in a subset of fibers (Figure 1C), confirming expression of peptidergic and non-peptidergic meningeal primary afferent neurons.
Figure 1. Anatomy of meningeal innervation. (A) Schematic of the injection strategy for AAV-mediated expression of GCaMP6s in trigeminal ganglion neurons without damaging the ipsilateral meninges. (B) Histological sections demonstrate GCaMP6s expression in a subset of neurons in the trigeminal ganglion. (C) Using immunohistochemistry, researchers found CGRP expression in 31.7-62.2% of dural GCaMP6s-labeled fibers, indicating that AAV transduces both peptidergic and non-peptidergic meningeal afferents. (Blaeser A S, et al., 2022)
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Our lab has been using AAV5-CAG-GCaMP6s for several months now, and the results have been outstanding. The consistency in gene expression across our experimental models has greatly enhanced our research efficiency.
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