Cat.No. : LV00951Z
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. | LV00951Z |
| Description | This lentivirus contains tdTOMATO under the control of CMV promoter and puromycin selection marker under the control of PGK promoter. |
| Target Gene | tdTOMATO |
| 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. |
TNS4 (Tensin 4 or Cten) is a putative oncogene in colorectal cancer (CRC) that plays a role in regulating cell adhesion, motility, invasion, and epithelial-mesenchymal transition (EMT). The effects of TNS4 knockdown on gefitinib chemosensitivity and epidermal growth factor receptor (EGFR) and Ras protein levels were also tested. In general, TNS4 knockdown increased cell proliferation in cell lines that generate dense spheroids. Addition of cancer-associated fibroblasts (CAFs) to spheroids supported CRC cell proliferation, whereas CAFs themselves did not proliferate but increased ECM degradation. TNS4 knockdown reduced adhesion and 3D invasiveness and disrupted EGFR signaling, resulting in increased sensitivity to gefitinib. In summary, in the 3D spheroid model, TNS4 inhibited cell proliferation and promoted cell invasion into the ECM, likely through adhesion to the ECM and stromal cells. TNS4 knockdown enhances sensitivity to the EGFR inhibitor gefitinib and may be helpful for CRC patients with Kirsten ras oncogene homozygous mutations.
Stable knockdown of TNS4 was achieved by lentiviral transduction of colorectal cell lines, which was confirmed by western blotting. Following stable knockdown of TNS4, transduction with TdTomato lentivirus allowed the researchers to track the growth of CRC cell populations in spheroids by detecting the expression of TdTomato fluorescent protein using a traditional fluorescent plate reader. The spheroid formation method used revealed different morphologies between CRC cell lines. While DLD1, HT29, and HCT116 formed compact spheroids with rounded uniform shapes, SW480 and SW620 produced aggregates, and LS1034 produced spheroids with irregular shapes.
Figure 1. A) Western blot and B) respective densitometry showed decreased TNS4 expression after stable knockdown of shTNS4 in DLD1 (62.3%), HT29 (39.4%), HCT116 (68.1%), SW480 (34.1%), and LS1034 (39.42%) cell lines compared with the loading control (β-Actin) and normalized to their shLUCs (luciferase control, 100%). (Raposo T P, Susanti S, Ilyas M., 2020)
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Using tdTOMATO Lentivirus, we achieved consistent results in our in vivo studies. It is a powerful tool for delivering genes to a wide range of tissues, enabling advanced research possibilities.
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