Cat.No. : LVIM013Z
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. | LVIM013Z |
| Description | Lentivirus expressing untagged Human HOXB8 under the control of CMV promoter, and puromycin resistance marker for mammalian cell selection. |
| Target Gene | HOXB8 |
| Species | Human |
| Application | Human HOXB8 lentivirus (CMV, Puro) is an important tool with a wide range of applications in cell biology, gene therapy, and cancer research. The following are detailed applications of HOXB8 lentiviral tools in various fields: 1. Cell immortalization: HOXB8 is a key homeobox-containing gene that has been shown to play an important role in cell immortalization. Gene introduction through HOXB8 lentivirus can enable primary cells (such as bone marrow-derived macrophages, fibroblasts, etc.) to grow in vitro for a long time without senescence. Such immortalized cells can provide consistency and stability for long-term biological studies. 2. Translational and cancer research: The expression of the HOXB8 gene is elevated in many cancers and is associated with tumor growth and invasiveness. Therefore, the HOXB8 lentiviral tool has been widely used in tumor research. By introducing HOXB8 into cell lines cultured in vitro, its specific mechanism of action and signaling pathways in cancer cells can be studied. 3. Gene function research: HOXB8 lentivirus can also be used to study its function in cell differentiation, development and morphogenesis. Using this viral vector, scientists can overexpress HOXB8 in different types of cells to observe its effects on gene expression profiles, cell behavior and morphology. 4. Gene therapy: Due to the important role of HOXB8 in cell proliferation and differentiation, its regulation may have potential for the treatment of certain diseases. By achieving proper expression regulation of HOXB8 lentivirus in specific cells or tissues, it is expected to develop new gene therapy methods to treat, for example, certain genetic diseases or degenerative diseases. 5. Drug screening: Cell immortalization makes the cell lines generated by HOXB8 lentivirus an ideal model for drug screening. These stable cell lines can be used for high-throughput screening to find new drugs targeting the HOXB8 pathway or discover its potential therapeutic targets. |
| 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. |
To present antigens to cognate T cells, dendritic cells (DCs) utilize the chemokine receptor CCR7 to enter lymph nodes directly from peripheral tissues via afferent lymphatics and through the floor of the subcapsular sinus into draining lymph nodes. Here, researchers combined the immortal proliferation capacity of conditionally Hoxb8-immortalized hematopoietic progenitor cells with CRISPR/Cas9 technology to create a powerful experimental system to study DC migration and function. Hematopoietic progenitor cells from the bone marrow of Cas9 transgenic mice were conditionally immortalized by lentiviral transduction to introduce a doxycycline-regulatable form of the transcription factor Hoxb8 (Cas9-Hoxb8 cells). The modified Cas9-Hoxb8 cells retained the potential to differentiate into myeloid cells in vitro, while GM-CSF-differentiated Cas9-Hoxb8 cells exhibited the classical phenotype of GM-CSF-differentiated bone marrow-derived dendritic cells. Finally, the researchers used two-photon microscopy to image Cas9-Hoxb8 dendritic cells expressing the genetic Ca2+ sensor GCaMP6S to observe chemokine-induced Ca2+ signaling in lymphoid-derived dendritic cells entering the lymph node parenchyma in real time.
Here, researchers used Hoxb8 lentiviral vectors for mouse hematopoietic cell immortalization. To validate vector-dependent immortalization of primitive hematopoietic stem and progenitor cells, lineage-negative cells were first transduced from C57BL/6J mice and then cultured in the presence of mSCF, huIL-11, huFlt3L, mIL-3, Dox, and Puro. Cells expressing the Hoxb8 construct were rescued from Puro treatment by day 15 post-transduction (pt), while non-transduced cells exhibited low cell numbers and viability (Figure 1A). To further utilize CRISPR/Cas9 technology for genome modification, researchers isolated lineage-negative cells from Cas9 mice and screened using the same strategy as C57BL/6J mice. Only gene-modified cells survived for more than 15 days (Figure 1B). These cells could be stably cultured for at least 16 weeks for further experiments and were named Cas9-Hoxb8 cells.
Figure 1. Immunoengineering of conditionally immortalized Hoxb8 cells from murine bone marrow. (Hammerschmidt S I, et al., 2018)
A: The cloning capacity for the transgene is approximately 3-4 kb for most vector formats.
A: Compared to conventional stable cell line constructionlentivirus exhibits a much higher positive clone rate with the target always co-existing with the selection marker. The cost labor. and time are substantially lower than transfection based stable cell line generation.
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This lentivirus expresses untagged human HOXB8 under the control of the CMV promoter and bears a mammalian cell-selected puromycin resistance marker, which not only saves time but makes colony selection easier.
The product can be stably integrated into the genome of host cells, independent of the cell cycle, and has good transduction efficiency.
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