Cat.No. : LVIM021Z
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. | LVIM021Z |
| Description | Lentivirus expressing untagged HPV16 E6-E7 under the control of CMV promoter, and puromycin resistance marker for mammalian cell selection. |
| Target Gene | E6-E7 |
| Species | HPV16 |
| Application | HPV16 E6-E7 lentivirus (CMV, Puro) is a powerful tool widely used in molecular biology and cell research for cell immortalization. Here are some specific applications: 1. Cell immortalization: Many primary cells have a limited lifespan and become senescent after a few population doublings. Introduction of HPV16 E6-E7 proteins via lentiviral transduction can extend the replicative capacity of these cells, resulting in immortalized cell lines. 2. Cancer research: Because the E6 and E7 oncoproteins can inactivate the p53 and pRB tumor suppressor proteins, respectively, transduction enables researchers to study the mechanisms of tumorigenesis and tumor progression. 3. Drug screening and development: Immortalized cells expressing these viral oncoproteins can be used to screen anticancer drugs, understand mechanisms of drug resistance, and identify potential therapeutic targets. 4. Gene editing and CRISPR: Immortalized cells provide an excellent system for genetic manipulation. They can be easily edited using the CRISPR/Cas9 system, allowing the creation of knockout, knockin, or other genetically modified cell lines for functional studies. 5. Virology and infectious disease research: Immortalized cells provide a model system for studying various aspects of the viral life cycle, replication, and host-virus interactions. |
| 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. |
High-risk human papillomavirus (HPV), especially HPV16, is closely associated with the development of human esophageal squamous cell carcinoma (ESCC). The studies here showed that ESCC cells stably transfected with HPV16 E6-E7 lentiviral vectors exhibited obvious cancer stem-like cell (CSCs) phenotypes, such as migration, invasion, sphere formation, high expression of p75NTR (a CSCs marker in ESCC), chemotherapy resistance, radioresistance, anti-apoptosis ability in vitro, and carcinogenicity in vivo. HPV16 E6-E7 induced activation of the PI3K/Akt signaling pathway, and this effect could be effectively inhibited by the specific PI3K inhibitor LY294002. It was also shown that inhibition of the PI3K/Akt signaling pathway by PI3K and Akt siRNA could reverse the induction of HPV16 E6-E7 on ESCC cells. In conclusion, these studies indicate that HPV16 E6-E7 promotes the CSC phenotype in ESCC cells by activating the PI3K/Akt signaling pathway. Targeting the PI3K/Akt signaling pathway in HPV16-positive tissues is a feasible treatment for ESCC patients.
Here, HPV16 E6-E7 protein was stably expressed in ESCC cells transfected with HPV16 E6-E7 lentiviral vectors, which were labeled as Eca109-psb and TE-1-psb cells, respectively. No HPV16 E6-E7 protein expression was observed in ESCC cells transfected with control vectors, which were labeled as Eca109-control and TE-1-control cells, respectively. The transfection efficiency was detected and verified by Western blotting (Figure 1A). After 72 h of experimental treatment of Eca109 and TE-1 cells, the transfection results of HPV16 E6-E7 lentiviral vectors were observed by fluorescence microscopy (Figure 1B).
Figure 1. HPV16 E6-E7 lentiviral vector stably transfected ESCC cells. (Xi R, et al., 2016)
A: For lentiviruses, they are measured in CFU/ml (colony-forming units per millilitre). Transduction with lentiviruses and retroviruses can cause the formation of colonies, which can be quantified for concentration.
A: For your reference, we recommend the following amount of virus-containing media for infection: 10-cm plate: 8-10 ml per plate; 6-well plate: 1 ml per well; 12-well plate: 0.5 ml per well; 24-well plate: 0.2 ml per well; This roughly reflects the surface area of each well or plate.
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High-titer lentivirus containing the HPV-16 E6/E7 gene that is well suited for cell immortalization.
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