Cat.No. : AD00239Z
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. | AD00239Z |
| Target Gene | PROX1 |
| Species | Human |
| Product Type | Adenoviral particle |
| Insert | PROX1 |
| 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. |
| Gene Name | PROX1 prospero homeobox 1 [ Homo sapiens ] |
| Gene Symbol | PROX1 |
| Gene ID | 5629 |
| UniProt ID | Q92786 |
| mRNA Refseq | NM_001270616.1 |
| Protein Refseq | NP_001257545.1 |
| Chromosome Location | 1q41 |
| Function | DBD domain binding; DNA binding; LBD domain binding; RNA polymerase II distal enhancer sequence-specific DNA binding transcription factor activity; ligand-dependent nuclear receptor binding; protein binding; sequence-specific DNA binding; sequence-specific DNA binding transcription factor activity; transcription corepressor activity; transcription regulatory region DNA binding; |
| MIM | 601546 |
Hepatocyte-like cells differentiated from human iPS cells (human iPS-HLCs) hold promise for drug development and research. However, recent hepatic characterization of human iPS-HLCs suggests that these cells resemble fetal hepatocytes more than adult hepatocytes. Here, researchers aimed to develop a method to enhance the hepatic function of human iPS-HLCs. Because gene expression levels of hepatic transcription factors (activating transcription factor 5 (ATF5), CCAAT/enhancer binding protein α (c/EBPα), and prospero homeobox protein 1 (PROX1)) are significantly higher in adult liver than in human iPS-HLCs and fetal liver, researchers expected that the hepatic function of human iPS-HLCs could be enhanced by adenoviral (Ad) vector-mediated transduction of ATF5, c/EBPα, and PROX1. ATF5, c/EBPα, and PROX1 transduction upregulated the gene expression levels of cytochrome P450 (CYP) 2C9, 2E1, α-1 antitrypsin, transthyretin, Na+/taurocholate cotransporting polypeptide, and uridine diphosphate glucuronosyltransferase 1A1, and the protein expression levels of CYP2C9 and CYP2E1. These results suggest that ATF5, c/EBPα, and PROX1 transduction can enhance the hepatic function of human iPS-HLCs.
Hepatic differentiation was performed according to the protocol shown in Figure 1A to generate the human iPS-HLCs. Since the expression levels of ATF5, c/EBPα, and PROX1 (three transcription factors: 3TF) in human iPS-HLCs and fetal livers are lower than those in adult livers (Figure 1B), it is expected that overexpression of 3TFs will enhance the hepatic function of human iPS-HLCs. To overexpress 3TFs in human iPS-HLCs, Ad vectors expressing ATF5 (Ad-ATF5), Ad vectors expressing c/EBPα (Ad-c/EBPα), and Ad vectors expressing PROX1 (Ad-PROX1) were used here. Human iPS-HLCs were transduced with Ad-LacZ at 3000 VP/cell, and LacZ expression in the cells was examined by X-Gal staining. Human iPS-HLCs transduced with Ad-LacZ uniformly expressed LacZ (Figure 1C). This result suggests that the Ad vectors used can efficiently overexpress transgenes in human iPS-HLCs. Next, human iPS-HLCs were transduced with Ad-ATF5, Ad-c/EBPα, and Ad-PROX1 at 1000 VP/cell/each Ad vector. Successful overexpression of 3TFs using Ad-3TFs was confirmed (Figure 1D).
Figure 1. Efficient Ad vector-mediated hepatic gene transfer into the human iPS cell-derived hepatocyte-like cells. (Nakamori D, et al., 2016)
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These PROX1 adenoviral particles performed exactly as expected. Transduction efficiency in our primary endothelial cells was very high, and PROX1 expression was stable and easily detectable. Very efficient!
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