Cat.No. : AAV00074Z
Titer: ≥1x10^12 GC/mL / ≥1x10^13 GC/mL Size: 30 ul/100 ul/500 ul/1 ml
Serotype: AAV Serotype 1 Storage: -80 ℃
| Cat. No. | AAV00074Z |
| Description | Synapsin-GFP AAV (Serotype 1) is the serotype 2/1 (with Capsid from AAV1 and ITR from AAV2) which express eGFP under the Synapsin promoter which restricts transgene expression only in neurons. |
| Reporter | GFP |
| Serotype | AAV Serotype 1 |
| Target Gene | Synapsin-GFP |
| Product Type | Adeno-associated virus |
| Application | 1. Determination of optimal MOI (multiplicity of infection), administration methods etc. 2. Detection of the infection efficiency of the AAV serotype against a specific cell type or tissue. 3. Using reporter genes to visualize the distribution and expression of AAV vectors in live animals, helping assess the biodistribution and persistence of gene delivery. |
| 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. |
mTOR signaling involves the mTORC1 and mTORC2 complexes, which critically regulate neurodevelopment and are implicated in various brain disorders. Here, researchers demonstrate direct, controlled inhibition of mTOR by Tanc2, an adaptor/scaffold protein with strong neurodevelopmental and psychiatric effects. While Tanc2-null mice exhibit embryonic lethality, Tanc2-haploinsufficient mice survive but exhibit hyperactivity of mTORC1/2 with concomitant synaptic and behavioral defects that are reversed by mTOR inhibition with rapamycin. Tanc2 interacts with and inhibits mTOR, which is inhibited by mTOR-activating serum or the rapid-acting antidepressant ketamine. Tanc2 and Deptor, also known to inhibit mTORC1/2, have minimal effects on neurodevelopment but clearly inhibit mTOR in both early and late neurons. Finally, Tanc2 inhibits mTORC1/2 in human neural progenitor cells and neurons. These results suggest that Tanc2 is a mTORC1/2 inhibitor that affects neural development.
Because the mTOR hyperactivity observed in Tanc2+/− pups (P14) could represent indirect changes resulting from the long-term absence of Tanc2, researchers generated a Tanc2 mutant mouse line carrying a floxed Tanc2 allele (Tanc2fl/fl) to create a conditional gene knockout (cKO). Local homozygous knockout of Tanc2 in the hippocampus of Tanc2fl/fl mice by injection of AAV1-Synapsin-Cre-eGFP and AAV1-Synapsin-eGFP (control) at P5 and analysis at P14, but not the injection at P19 and analysis at P28, induced hyper-phosphorylation of S6 (S235/236), 4E-BP (T37/46), Akt (S473), GSK3β (S9), and mTOR (S2248) (Figure 1e–h), indicative of mTORC1 and mTORC2 hyperactivity. These results indicate that Tanc2 deletion leads to mTORC1/2 hyperactivity at the pup (P7-14) stage, but not at the juvenile (P21-28) or adult (~P52) stages.
Figure 1. e, f Cre-dependent acute hippocampal Tanc2 deletion during P5–14 resulted in increased phosphorylation of S6 (S235/236), 4E-BP (T37/46), Akt (S473), GSK3β (S9), and mTOR (S2448), indicating hyperactivity of mTORC1 and mTORC2. AAV1-Synapsin-Cre-eGFP and AAV1-Synapsin-eGFP (control) were injected in parallel into the hippocampus (CA3 region) of Tanc2fl/fl mice bilaterally (eGFP expression is indicated in green). g, h Cre-dependent acute hippocampal (CA3 region) Tanc2 deletion during P19–28 does not increase phosphorylation of S6 (S235/236), 4E-BP (T37/46), Akt (S473), GSK3β (S9), or mTOR (S2448), indicating the absence of mTORC1 and mTORC2 hyperactivity. (Kim S G, et al., 2021)
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