Cat.No. : LVIM001Z
Titer: ≥1*10^7 TU/mL / ≥1*10^8 TU/mL / ≥1*10^9 TU/mL Size: 100 ul/500 ul/1 mL
Storage: -80℃ Shipping: Frozen on dry ice
| Cat. No. | LVIM001Z |
| Description | Lentivirus expressing untagged SV40 Small T and Large T Antigen under the control of CMV promoter, and puromycin resistance marker for mammalian cell selection. |
| Target Gene | Small/Large T Antigen |
| Species | SV40 |
| Application | SV40 small/large T antigen lentiviruses (CMV, Puro) are commonly used research tools in molecular and cellular biology. Here are some key applications: Cell immortalization: SV40 large T antigen can extend the lifespan of primary cells by interfering with cell cycle regulatory proteins. This allows researchers to create immortalized cell lines that can proliferate indefinitely, facilitating long-term studies and experiments. Gene expression studies: Lentiviral vectors are often used to deliver and express SV40 T antigen in target cells. Using the CMV promoter to drive transgene expression, you can achieve high levels of expression, which makes it useful for overexpression studies. Cancer research: By immortalizing cells and manipulating their growth, researchers can study transformations and pathways that lead to carcinogenesis. SV40 T antigen has been implicated in disrupting tumor suppressor pathways such as p53 and Rb, making this tool valuable for cancer research. Protein interaction studies: SV40 T antigen interacts with a variety of host proteins. By studying these interactions, researchers can gain insight into cellular mechanisms and the regulation of the cell cycle and apoptosis. Stable cell line generation: The puromycin resistance gene (Puro) allows selection of successfully transduced cells, ensuring that only cells expressing the SV40 T antigen survive in the presence of puromycin. This is critical for generating stable cell lines for downstream applications. |
| Titer | Varies lot by lot, for example, ≥1*10^7 TU/mL, ≥1*10^8 TU/mL, ≥1*10^9 TU/mL etc. |
| Size | Varies lot by lot, for example, 100 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 lentivirus particles by optimizing and standardizing production protocols and performing stringent quality control (QC). The specific QC experiments performed vary between lentivirus particle lots. | |
| Mycoplasma | Creative Biogene routinely tests for mycoplasma contamination using a mycoplasma detection kit. Cell lines are maintained for approximately 20 passages before being discarded and replaced with a new vial of early passage cells. Approximately 2 weeks after thawing, cell culture supernatants are tested for mycoplasma contamination. Creative Biogene ensures that lentiviral products are free of mycoplasma contamination. |
| Purity | Creative Biogene evaluates the level of impurities, such as residual host cell DNA or proteins, in prepared lentiviral vectors to ensure they meet quality standards. |
| Sterility | The lentiviral samples were inoculated into cell culture medium for about 5 days and the growth of bacteria and fungi was tested. Creative Biogene ensures that the lentiviral products are free of microbial contamination. |
| Transducibility | Upon requirement, Creative Biogene can perform in vitro or in vivo transduction assays to evaluate the ability of lentivirus to deliver genetic material into target cells, and assess gene expression and functional activities. |
| Proviral Identity Confirmation | All Creative Biogene lentiviral vectors are confirmed to have correctly integrated provirus using PCR. This test involves transducing cells with serial dilutions of the lentiviral vector, harvesting the cells a few days later, and isolating genomic DNA. This DNA is then used as a template to amplify a portion of the expected lentiviral insert. |
The hyperinsulinemic hypoglycemic isoform of glucokinase (GCK-HH) is caused by activating mutations in glucokinase (GCK) and has been shown to increase β-cell death. However, the mechanism of β-cell death in GCK-HH remains poorly understood. Here, researchers expressed the GCK-HH V91L GCK mutant in INS-1 832/13 cells to determine the effects of the mutation on β-cell viability and the mechanism of β-cell death. They found that expression of the V91L GCK mutant in INS-1 832/13 cells resulted in a rapid loss of cell viability that was glucose concentration-dependent. Under 11 mM D-glucose, INS-1 832/13 cells expressing V91L GCK had increased cell permeability without significant increases in Annexin V staining or caspase 3/7 activation, suggesting that these cells undergo cell death primarily through necrosis. Overexpression of the SV40 large T antigen, which inhibits the p53 pathway, did not affect V91L GCK-induced cell death. The researchers also found that non-phosphorylatable L-glucose did not induce rapid cell death. These data suggest that the GCK V91L mutant induces rapid necrosis in INS-1 cells by accelerating glucose phosphorylation, ATP depletion, and increasing cell permeability.
To determine whether p53 activation is involved in D-glucose-induced cell death in INS-1 832/13 GCK V91L cells, the researchers expressed simian virus 40 (SV40) small and large T antigens by lentiviral transduction. Expression of SV40 large T antigen in INS-1 832/13 cells after lentiviral vector transduction was confirmed by immunoblotting (Figure 1A). Transduction of SV40 Small/Large T Antigen Lentivirus did not rescue INS-1 832/13 GCK V91L cells from D-glucose-induced cell death (Figure 1B), indicating that p53 activation is not the primary mechanism of cell death. The study also showed that glucose phosphorylation, but not glucose metabolism, is required for glucose-induced cell death in INS-1 832/13 GCK V91L cells (Figure 1C). Furthermore, incubation of INS-1 832/13 GCK V91L cells with 11 mM D-glucose or 2-DG resulted in a loss of more than 90% of ATP content within 1 h, whereas incubation with 11 mM L-glucose did not result in changes in cellular ATP content (Figure 1D). These results suggest that overactive V91L GCK induces cell death by depleting cellular ATP and increasing cell permeability.
Figure 1. Glucose phosphorylation is essential for D-glucose-induced death of INS-1 832/13 GCK V91L cells. (Lu B, et al., 2019)
A: SV40 large T-antigen causes cell immortalization and allow to produce high-titer lentivirus in your cell line. It was natively expressed without any tags.
A: The large and small tumor antigens (T antigens) are the major regulatory proteins encoded by SV40. Large T antigen is responsible for both viral and cellular transcriptional regulation, virion assembly, viral DNA replication, and alteration of the cell cycle.
A: You can observe transduction efficiency from 48 hours up to 5 days after infection.
If your question is not addressed through these resources, you can fill out the online form below and we will answer your question as soon as possible.
Ready-to-use lentiviral particles for the transduction of a variety of mammalian cells includingdifficult-to-transfect, primary, stem and non-dividing cells.
High titer product.Reliable and commonly used. Often used to immortalize epithelial cells.
Write a review of your use of Biogene products and services in your research. Your review can help your fellow researchers make informed purchasing decisions.
Our promise to you:
Guaranteed product quality, expert customer support.
24x7 CUSTOMER SERVICE
CONTACT US TO ORDER
Copyright © Creative Biogene. All rights reserved.
