Cat.No. : AD00195Z
Titer: ≥1x10^10 IFU/mL / ≥1x10^11 IFU/mL / ≥1x10^11 VP/mL / ≥1x10^12 VP/mL Size: 100 ul/500 ul/1 mL
Storage: -80℃ Shipping: Frozen on dry ice
| Cat. No. | AD00195Z |
| Target Gene | KLF4 |
| Species | Human |
| Product Type | Adenoviral particle |
| Insert | KLF4 |
| Titer | Varies lot by lot, for example, ≥1x10^10 IFU/mL, ≥1x10^11 IFU/mL, ≥1x10^11 VP/mL etc. |
| Size | Varies lot by lot, for example, 250 ul, 500 ul, 1 mL etc. |
| Storage | Store at -80℃. Avoid multiple freeze/thaw cycles. |
| Shipping | Frozen on dry ice |
| Creative Biogene ensures high-quality adenovirus particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between adenovirus particle lots. | |
| Endotoxin | Endotoxins, primarily derived from Gram-negative bacteria, can trigger adverse immune responses. Endotoxin contamination is a significant concern in adenovirus production, especially for applications in animal studies and gene therapy. Creative Biogene utilizes rigorous endotoxin detection methods to monitor the endotoxin level in our produced adenovirus particles to ensure regulatory compliance. |
| Sterility | Creative Biogene ensures that adenovirus products are free of any bacterial, fungal and other microbial contamination. |
| Ad5 E1 Detection | All Creative Biogene adenoviruses are PCR tested to ensure that there are no detectable E1 sequences in the particles, which could be from revertants or external E1 contamination. |
| RCA Assays | Adenovirus products originating at Creative Biogene are guaranteed to have undetectable replication-competent adenovirus (RCA). This quality control measure is important because there is always the possibility of wild-type contamination due to revertants or environmental sources. |
| PFU Titering | All purified adenovirus preparations are tested for infectious titer. Creative Biogene's PFU test takes a few days longer but counts true plaques in HEK cells rather than estimating PFU titers via IHC staining or TCI50 of infected cells. |
Endometrial angiogenesis is essential to maintain good endometrial receptivity. Krüppel-like factor 4 (KLF4) is a transcription factor that is essential for regulating angiogenesis. Here, researchers found that vascular endothelial growth factor A (VEGFA) can form a positive feedback loop with KLF4 to promote proliferation and migration of human endometrial microvascular endothelial cells (HEMECs) and inhibit apoptosis. General control nonrepressible 5 (GCN5) also has a time-dependent response to VEGFA and participates in the KLF4-VEGFA loop. In addition, researchers found that GCN5 is a succinyltransferase that regulates the succinylation of histones and non-histone proteins. GCN5 interacts with KLF4 and is recruited to the KLF4 binding site of the VEGFA promoter to succinylate H3K79, thereby epigenetically initiating gene transcription. For non-histone proteins, GCN5 succinylated KLF4, which was activated by ERK signaling in HEMECs treated with VEGFA, thereby increasing its transcriptional activity. These results suggest that the KLF4-VEGFA positive feedback loop is epigenetically regulated, which contributes to endometrial angiogenesis.
In vitro, HEMECs were co-treated with KLF4 overexpression adenovirus (Ad-KLF4) or VEGFA. KLF4 overexpression further increased PCNA and MMP-9 and decreased Caspase 3 expression by VEGFA (Figure 1A). In contrast, when HEMECs were infected with three KLF4-specific shRNA (sh-KLF4) adenoviruses to block endogenous KLF4 expression (sh-KLF4 2 and sh-KLF4 3 had high knockdown efficiency), VEGFA-induced PCNA and MMP-9 expression and VEGFA-reduced Caspase 3 expression were abolished (Figure 1B-C). In addition, cell immunofluorescence assays showed that KLF4 overexpression promoted Ki67 expression, and the number of Ki67-positive cells decreased after KLF4 knockdown (Figures 1D and 1F). The scratch-wound assay showed that the KLF4 overexpression and VEGFA co-treatment group had stronger migration ability than the Ad group, while the KLF4 knockdown and VEGFA co-treatment group had weaker migration ability than the VEGFA group (Figure 1E and 1G). These results indicate that KLF4 is essential for inducing endometrial angiogenesis by regulating proliferation, migration, and apoptosis caused by VEGFA signaling.
Figure 1. KLF4 mediates VEGFA-induced endometrial angiogenesis. (Cao C, et al., 2022)
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