Cat.No. : AAV00322Z
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV serotype Retrograde Storage: -80 ℃
| Cat. No. | AAV00322Z |
| Description | Prepackaged AAV particles in serotype retrograde containing Cre recombinase gene under the control of CAG promoter. |
| Serotype | AAV serotype Retrograde |
| Target Gene | Cre |
| 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. |
Marked deficits in glucose availability, or glucoprivation, elicit organism-wide counter-regulatory responses whose purpose is to restore glucose homeostasis. However, although catecholamine neurons of the ventrolateral medulla (VLMCA) are thought to coordinate these responses, the circuits and cellular mechanisms underlying specific counterregulatory responses are largely unknown. Here, researchers combine anatomical, imaging, optogenetic, and behavioral approaches to explore the circuit mechanisms by which VLMCA neurons coordinate food-seeking behavior induced by glucose deprivation. Using these methods, they found that VLMCA neurons form functional connections with nucleus accumbens (NAc) projection neurons in the posterior paraventricular nucleus of the thalamus (pPVT). Importantly, optogenetic manipulations demonstrate that while activating VLMCA projections to the pPVT is sufficient to elicit robust feeding behavior in well fed mice, inhibition of VLMCA-pPVT communication significantly impairs glucoprivation-induced feeding while leaving other major counterregulatory responses intact. Taken together, these findings indicate that the VLMCA-pPVT-NAc pathway is a previously overlooked node that selectively controls glucoprivation-induced food seeking.
Here, to probe whether VLMCA projections form functional connections with PVT-NAc neurons, researchers optogenetically stimulated VLMCA-PVT projections (using a red-shifted ChR2 variant; ChrimsonR) and imaged calcium responses in NAc-projecting pPVT neurons using fiber photometry in awake animals (Figure 1a-c). To achieve selective expression of the genetically encoded calcium sensor GCaMP6s in NAc-projecting neurons of the pPVT, they injected TH-Cre mice bilaterally into the NAc with a viral vector driving retrograde expression of Cre recombinase (AAV(retro)-CAG-Cre) followed by a vector encoding Cre-dependent GCaMP6s in the pPVT. Interestingly, optogenetic activation of VLMCA inputs significantly increased the fluorescence of the genetically encoded calcium sensor GCaMP6s in NAc-projecting pPVT neurons (Figure 1d-g). Notably, this effect was dependent on ChrimsonR expression in VLMCA neurons (Figure 1f, g).
Figure 1. Photostimulation of VLMCA axonal inputs to pPVT increases the activity of pPVT–NAc neurons. (Sofia Beas B, et al., 2020)
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