Cat.No. : LVIM006Z
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. | LVIM006Z |
| Description | Lentivirus expressing untagged Human TERT under the control of CMV promoter, and puromycin resistance marker for mammalian cell selection. |
| Target Gene | TERT |
| Species | Human |
| Application | The human TERT lentivirus (CMV, Puro) is a viral vector used primarily in research and biotech applications. Here are some of the main applications: Cell Immortality: The human telomerase reverse transcriptase (hTERT) component is responsible for maintaining telomere length, allowing cells to avoid senescence and continue to proliferate. Introducing hTERT into primary cells using this lentivirus can immortalize the cells, making them easier to maintain and study over longer periods of time. Cancer Research: Since telomerase activity is upregulated in many cancers, the role of telomerase in cancer progression can be studied using this lentivirus. By introducing hTERT into various cell lines, researchers can better understand its effects on cell behavior and tumorigenesis. Drug Screening: Immortalized cell lines generated with hTERT can be used for high-throughput screening of drug compounds. They provide a consistent and renewable source of cells for testing the effects of drugs on cell growth, survival, and telomerase activity. Human Disease Models: Since telomerase plays a role in a variety of diseases beyond cancer, creating cell models with altered telomerase activity could help study other telomere-related diseases such as dyskeratosis congenita or idiopathic pulmonary fibrosis. |
| 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. |
Human exfoliated deciduous tooth stem cells (SHEDs) have recently attracted attention as a novel source of multipotent stem cells. However, their application is limited due to replicative senescence in vitro. Ectopic expression of telomerase reverse transcriptase (TERT) is a promising strategy to overcome this replicative senescence. Using lentiviral transduction with a puromycin selection marker, TERT expression was stably restored in SHEDs isolated from healthy children aged 6-8 years (TERT-SHEDs). The study showed that lentiviral transduction induced stable TERT expression even in SHEDs at passage 40. TERT-SHEDs showed robust proliferation capacity and low concentrations of β-galactosidase. Although they had some different biomarkers from early-passage SHEDs, late-passage TERT-SHEDs showed multi-lineage differentiation similar to early-passage TERTs. In addition, late-passage TERT-SHEDs had normal karyotypes, no soft agar colony formation, and no tumor formation in nude mice. These data suggest that TERT-immortalized SHEDs may be a promising resource for stem cell therapy.
The multidirectional differentiation potential of SHED was measured by differentiation induction assay. SHED P4 could differentiate into osteogenic, adipogenic, and chondrogenic lineages, with positive staining by Alizarin Red S, Oil Red O, and Toluidine Blue, respectively (Figure 1a-c). TERT-SHED P20 showed similar differentiation ability to SHED P4 (Figure 1e-g). In addition, real-time PCR analysis confirmed that TERT-SHED P20 and SHED P4 had similar expression of osteogenic (ALP and BSP), adipogenic (LPL and PPAR-γ), and chondrogenic (ACAN and COL2A1) differentiation markers (Figure 1d and h).
Figure 1. Multilineage differentiation assay of SHED P4 and TERT-SHED P20. (Yin Z, et al., 2016)
β-GAL activity at pH 6 is a known characteristic of senescent cells, whereas this activity was not found in pre-senescent, quiescent, or immortalized cells. We first detected the senescence marker β-GAL in SHED and TERT-SHED at different passages. As shown in Figure 2a, the concentration of β-GAL in passage 20 SHED (SHED P20) was 120 times higher than that in SHED P4, but the concentration of β-GAL in passage 40 TERT-SHED (TERT-SHED P40) was still at a very low level. The proliferation capacity of SHED was detected by CCK-8 assay. As shown in Figure 2b, the proliferation capacity of late passage (P20) SHED was significantly decreased, while the proliferation potential of TERT-SHED P40 was similar to that of early passage (P4) SHED.
Figure 2. Comparison of β-GAL expression and proliferation ability between SHED and TERT-SHED of different passages. (Yin Z, et al., 2016)
A: Telomerase reverse transcriptase (TERT, or hTERT in human) is a catalytic subunit of the enzyme telomerase, Ectopic expression of telomerase in telomerase-silent cells is sufficient to overcome senescence and to extend cellular lifespan, and has been used for cell immortalization. This lentivirus expresses hTERT under the control of CMV promoter and has puromycin as a selection marker.
A: It depends on the purpose of the experiment. Higher titer of lentivirus should be used for in vivo experiment compared to in vitro experiment. Though lentiviral vector is highly efficient in transduction, the transduction efficiency of lentivirus depends heavily on the cell type to be transduced. Pilot experiment is highly recommended to determine how efficient the lentivirus on your target cells.
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.
Highly reproducible and controllable expression delivery method.
The pre-made lentiviral particles provide a ready-to-use easy delivery method that avoids the often troublesome lentivector contraction and lentiviral virus production.
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.
