EF1α-FLuc-IRES-Puro Lentivirus

Name: EF1α-FLuc-IRES-Puro Lentivirus
SKU: LV00996Z

Cat.No. : LV00996Z

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

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Lentivirus Particle Information

Quality Control

Cat. No.	LV00996Z
Description	This lentivirus contains Firefly luciferase reporter gene and IRES-mediated puromycin resistance gene under EF1a promoter.
Target Gene	Luciferase
Titer	Varies lot by lot, for example, ≥110^7 TU/mL, ≥110^8 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.

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Unlike free viruses (e.g. herpes simplex virus, adenovirus), retroviruses have the ability to integrate genetic material into the host genome, allowing for stable, long-term expression of transgenes. Therefore, simple retroviruses (so-called gamma or oncoretroviruses) play an important role in contemporary biomedicine. However, the inability of simple retroviruses to infect non-dividing cells and the risk of inducing cancer limit their application. The use of lentiviruses can circumvent these shortcomings of simple retroviruses. Lentiviruses are a class of complex retroviruses that contain additional regulatory and auxiliary genes in addition to the conventional genes of simple retroviruses such as gag, pol and env. Unlike simple retroviruses, lentiviruses can infect both dividing and non-dividing cells, greatly increasing the range of cells to which genes can be transferred. Lentiviruses tend to integrate into intronic regions of chromosomes (expressed genes are rich in introns), while simple retroviruses prefer promoters and regulatory regions. Widely used lentiviral vectors are engineered from human immunodeficiency virus type 1 (HIV-1).

HIV-1 virus particles are roughly spherical, about 100 nm in diameter, and are covered with a lipid bilayer membrane derived from the host cell during the budding process. The surface of the envelope carries many small glycoprotein protrusions. The virus recognizes new host cells through these glycoproteins. The matrix protein under the lipid layer covers the inner surface of the viral envelope to form a protective shell. Inside the virus particle is a conical capsid assembled by structural proteins, which contains the viral genome. Two single-stranded RNA genomes with a length of 9.2Kb, nucleocapsid protein, reverse transcriptase, and integrase are co-loaded in the capsid.

Radiation therapy for head and neck cancer often damages the salivary glands and oral mucosa, severely affecting patients' quality of life. FLASH proton radiotherapy (F-PRT) has previously been demonstrated in multiple tissues to reduce normal tissue toxicity while maintaining tumor control compared with standard proton radiotherapy (S-PRT). However, its potential to improve radiation-induced salivary gland dysfunction and oral mucositis and control the growth of orthotopic head and neck tumors has not been reported. Mice treated with single or fractionated doses of F-PRT had significantly higher survival rates than mice irradiated with S-PRT. F-PRT-treated mice had improved salivary flow. S-PRT-irradiated mice had increased fibrosis in the tongue epithelium. F-PRT significantly improved overall survival of mice with orthotopic tumors compared with mice treated with S-PRT. F-PRT demonstrated the ability to reduce radiation-induced normal tissue toxicity without compromising tumor control, suggesting that this modality may be useful in the clinical management of patients with head and neck cancer.

To evaluate the comparative efficacy of F-PRT versus S-PRT in a more relevant potential clinical scenario, orthotopic tongue tumors were generated using the MOC2 HNSCC cell line transfected with firefly luciferase and injected into the tongue of syngeneic C57Bl/6 mice. Here, lentiviral-mediated transfection of MOC2 cells was performed using EF1α-FLuc-IRES-Puro Lentivirus, and transfected cells were selected using the antibiotic puromycin (1 μg/mL). Orthotopic head and neck tumors were generated by injecting 5 X 10⁴ MOC2-luc cells (suspended in 30 μL PBS) directly into the lateral tongue. Following tumor implantation and dissemination, a dose escalation study of 14, 16, and 18 Gy PRT was performed (Figure 1A), and survival was studied at 90 days. The Kaplan-Meier plot (Figure 1B) showed that the overall survival of mice irradiated with F-PRT was significantly improved compared with S-PRT at dose levels of 14 Gy (P = 0.031), 16 Gy (P = 0.009), and 18 Gy (P = 0.032).

Figure 1. F-PRT increases overall survival in an orthotopic head and neck tumor model. (Chowdhury P, et al., 2024)

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