Cat.No. : AAV00154Z
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. | AAV00154Z |
| Description | Self-complementary AAV serotype 2 particles contain Cre recombinase under the control of CMV promoter. |
| Serotype | AAV Serotype 2 |
| 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. |
Primary sensory axons in adult mammals fail to regenerate after spinal cord injury (SCI), in part due to insufficient intrinsic growth potential. Previous studies have shown that constitutive activation of B-RAF (rapidly accelerating fibrosarcoma kinase) significantly promotes axonal regeneration after dorsal root and optic nerve injury. The regrowth was further enhanced by complementary deletion of PTEN (phosphatase and tensin homolog). Here, researchers examined whether simultaneous activation of B-RAF and deletion of PTEN would promote dorsal column axon regeneration after SCI. Notably, selective genetic targeting of B-RAF and PTEN in DRG neurons in adult mice enables many DC axons to enter, cross, and grow beyond the lesion site after SCI. Co-targeting of B-RAF and PTEN promotes more robust DC regeneration compared to preconditioned lesions, thus enhancing regeneration triggered by B-RAF/PTEN. Furthermore, post-injury targeting of B-RAF and PTEN enhances DC axon regeneration. These results demonstrate that simultaneous targeting of B-RAF and PTEN effectively enhances the intrinsic growth potential of dendrites after spinal cord injury and may therefore contribute to the development of a new strategy to promote robust long-distance regeneration of primary sensory axons.
Here, researchers injected scAAV2-Cre into L4 and L5 DRGs immediately after SCI in TdTom+/+ control and kaBRAF/PTEN/TdTom+/+ mice (Figure 1A). Lesion areas in TdTom+/+ controls were 0.056–0.120 mm2, while lesions in kaBRAF/PTEN/TdTom+/+ mice were 0.067–0.168 mm2 (Figure 1B). Quantification of lesion size showed no difference in lesion area between TdTom+/+ and kaBRAF/PTEN/TdTom+/+ groups (Figure 1B). Three weeks after SCI in TdTom+/+ mice, most DC axons terminated at the astrocyte border caudal to the injury site. DC axons were rarely observed in the lesion epicenter (Figure 1C-C'). The studies here demonstrate that scAAV2-Cre predominantly labels large-diameter sensory neurons and that simultaneous B-RAF- and PTEN-targeted treatment immediately after SCI enhances DC axon regeneration, albeit at a weaker level compared with animals transduced 2 weeks before injury (Figure 1C–E).
Figure 1. Co-targeting of B-RAF and PTEN immediately after SCI promotes DC axon regeneration. (Noristani H N, et al., 2022)
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We have used the scAAV2-Cre vector in numerous in vivo and in vitro studies, and it has consistently delivered reliable results.
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