Cat.No. : AAV00504Z
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. | AAV00504Z |
| Description | AAV serotype 5 particles express Cre recombinase under the control of human Synapsin promoter for neuronal specific expression. |
| Serotype | AAV Serotype 5 |
| 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. |
Clock neurons within the mammalian suprachiasmatic nucleus (SCN) encode circadian time using an interlocking transcription-translation feedback loop (TTFL) to drive rhythmic gene expression. However, the contribution of transcription factors beyond the circadian TTFL to SCN function remains unclear. Here, researchers report that the stem and progenitor transcription factor, sex-determining region Y-box 2 (SOX2), is expressed in adult SCN neurons and positively regulates transcription of the core clock gene Period2. Mice lacking SOX2 selectively in SCN neurons exhibit imprecise, poorly consolidated behavioral rhythms that fail to effectively synchronize to the ambient light cycle and are highly susceptible to sustained light-induced arrhythmias. RNA sequencing reveals that Sox2 deficiency alters the SCN transcriptome, reducing expression of core clock genes and neuropeptide receptor systems. By defining the transcriptional landscape within SCN neurons, SOX2 enables the generation of stable, trainable circadian rhythms that accurately reflect environmental time.
To determine whether the effects of Sox2 disruption on PER2 expression were intrinsic to the fully developed SCN, researchers used adeno-associated virus (AAV)-mediated delivery of cre recombinase to ablate the Sox2 gene in neurons in SCN tissue explants prepared from Sox2fl/fl;Per2Luc/+ pups at postnatal day (P) 12 (Figure 1K) and monitored PER2::LUCIFERASE rhythms in culture. As most developmental milestones of the mouse SCN have been reached by ∼P10, this approach could avoid any potential effects of Sox2 ablation on SCN development. The amplitude of PER2::LUC rhythms was significantly reduced in cultures transduced with AAV5-hSyn-Cre compared with SCN explants transduced with control AAV5-human synaptobrevin (hSyn)-TurboRFP (Figures 1L and 1M). Collectively, these data suggest that SOX2 is important for the generation of high-amplitude PER2 rhythms within the SCN.
Figure 1. (K) SOX2 (green) and RFP (magenta) expression in Sox2fl/fl;Per2Luc/+ SCN tissue explants transduced with AAV5-hSyn-TurboRFP or AAV5-hSyn-Cre. (L) Bioluminescence traces from Sox2fl/fl;Per2Luc/+ SCN tissue explants that had been transduced with AAV5-hSyn-TurboRFP or AAV5-hSyn-Cre on day 5 after initial plating. (M) Relative amplitude of bioluminescence rhythms of transduced Sox2fl/fl;Per2Luc/+ SCN tissue explants after medium change. (Cheng A H, et al., 2019)
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