Cat.No. : AAV00082Z
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV Serotype 2 Storage: -80 ℃
| Cat. No. | AAV00082Z |
| Description | CAG-ChR2-YFP AAV (Serotype 2) is the serotype 2 AAV which express Channelrhodopsin-2 under CAG promoter with YFP tag. Channelrhodopsins are a subfamily of opsin proteins that function as light-gated ion channels and very useful for many bioengineering and neuroscience applications such as photostimulation of neurons for probing of neural circuits. Using a Yellow Fluorescent Protein (YFP) tagged ChR2, light-stimulated axons and synapses can be identified in intact brain tissue. This is useful to study the molecular events during the induction of synaptic plasticity. ChR2 has also been used to map long-range connections from one side of the brain to the other, and to map the spatial location of specific inputs on the dendritic tree of individual neurons. |
| Serotype | AAV Serotype 2 |
| Target Gene | CAG-ChR2-YFP |
| Product Type | Adeno-associated virus |
| 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. |
Although the medial prefrontal cortex (mPFC) is traditionally defined as reciprocal connections with the medial thalamic nucleus (MD), the nature of information transfer between the MD and mPFC is poorly understood. In the sensory thalamocortical pathway, thalamic recruitment of feedforward inhibition mediated by fast-firing, putatively parvalbumin (PV)-expressing interneurons is a key feature that enables cortical neurons to represent sensory stimuli with high temporal fidelity. Here, researchers show in mice that inputs from the MD drive bisynaptic feedforward inhibition in the dorsal anterior cingulate cortex (dACC) subregion of the mPFC. In particular, they demonstrate that axons from MD neurons directly synapse onto and excite PV interneurons, which in turn mediate feedforward inhibition of layer 3 pyramidal neurons in the dACC. This feedforward inhibition in the dACC restricts the temporal window in which pyramidal neurons can integrate excitatory synaptic inputs and fire action potentials, but in a manner that allows for greater flexibility than in the sensory cortex. These findings provide a foundation for understanding the role of MD-PFC circuit function in cognition.
To characterize the functional connectivity between the MD and dACC, the researchers injected the MD with AAV-CAG-ChR2-YFP, which expresses the light-sensitive cation channelrhodopsin-2 (ChR2) tagged with the yellow fluorescent protein (YFP) (Figure 1A, B). ChR2-YFP-expressing axon fibers originated from the MD and innervated different layers (L) of the dACC, including L1, L3, and L5, with the densest innervation in L3 (Figure 1B). The researchers stimulated these axons in acute slices using blue light pulses (0.5 ms) while recording EPSCs and IPSCs from L3 PNs in the dACC using the whole-cell patch clamp technique (Figure 1C). Photostimulation of MD axons reliably elicited both EPSCs and IPSCs in all L3 PNs recorded (Figure 1D), with IPSCs having a longer onset latency than EPSCs (Figure 1E). Application of the GABAA receptor antagonist picrotoxin or the AMPA/kainate receptor antagonist CNQX blocked IPSCs, indicating that they are polysynaptic inhibitory currents.
Figure 1. Inputs from MD drive excitation and inhibition onto layer 3 dACC pyramidal neurons. (Delevich K, et al., 2015)
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I am thoroughly impressed with the transduction efficiency of the CAG-ChR2-YFP AAV2. Even in difficult-to-transduce cell types, this product delivers robust and uniform expression of ChR2-YFP.
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