EF1α-tdTOMATO Lentivirus

Name: EF1α-tdTOMATO Lentivirus
SKU: LV00976Z

Cat.No. : LV00976Z

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.	LV00976Z
Description	This lentivirus contains tdTOMATO under the control of EF1α promoter.
Target Gene	tdTOMATO
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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Lentiviral vector technology is one of the effective tools for cultivating transgenic animal models. After the target gene is transferred into the host cell, it can be efficiently expressed in the host through reverse transcription, integration, etc., so as to obtain the required individuals with the target gene. Due to its simple operation and good adaptability to host cells, the lentiviral system has a good application prospect in transgenic animals. In many cases, the phenotypes of these mutant animals are similar to the clinical symptoms of a certain human disease, making these mutant animals an ideal model for human diseases. The disease phenotypes of genetically engineered animals obtained by transgenic and gene targeting can remain highly stable in inbred strains. Compared with models prepared by changing nutritional conditions, drug effects or surgical approaches, these genetically modified animal models have better consistency and stability, and can more realistically reflect the pathological processes and molecular changes of human diseases.

Numerous studies have shown that many genetically engineered animals (such as genetically engineered mice and miniature pigs) have become ideal disease animal models for studying major human diseases, including emerging and recurrent infectious diseases, tumors, cardiovascular diseases, senile diseases, mental diseases and genetic diseases. They play an important role in analyzing the pathogenic mechanisms and etiology of human diseases, animal and human behavior, the interaction between organisms and the environment, answering the susceptibility of specific populations to certain diseases, and developing new and effective preventive and therapeutic drugs.

The mammary glands of pigs shares many functional and morphological similarities with the human breast, raising their potential for use in studying the mechanisms underlying normal mammary function and breast carcinogenesis. Here, researchers sought to establish a model for efficient manipulation and transformation of porcine mammary epithelial cells (pMECs) in vitro and tumor growth in vivo. The study demonstrated that lentiviruses could effectively transduce pMECs in vitro and in vivo. It was further determined that lentiviruses could be used for oncogenic transformation of pMECs in vitro to generate mammary tumors in vivo. Oncogenic transformation in vitro was confirmed by anchorage-independent growth, increased cell proliferation, and expression of CDKN2A, cyclin A2, and p53, and decreased phosphorylation of Rb. Furthermore, Tag-transformed CD140a- and CD140a-CD49f + pMECs developed site-specific tumors with distinct histopathologies in vivo.

Here, researchers compared CMV, EF1α, and PGK promoters in lentivirally transduced pMECs and determined that EF1α was the most effective in vitro. Genomic DNA analysis showed that polybrene increased the incorporation of lentiviruses injected intraductally by 24-fold. The researchers also examined whether pMECs transduced in vitro would develop into epithelial structures after transplantation into donor pigs. After cell expansion and mounting 8 days after transduction, fluorescent TDLU outgrowths were generated in cells transduced with EF1α-tdTomato lentivirus (1 of 8 mammary glands) and CMV-tdTomato lentivirus (2 of 8) (Figure 1a), but not in PGK-tdTomato transducers or control glands, although tdTomato was detected in 2 of 6 glands transplanted with PGK-tdTomato pMECs (Figure 1b). Expansion of cells 24 h after transduction followed by mounting resulted in detection of genomic tdTomato in >50% of glands injected with CMV-tdTomato pMEC, EF1α-tdTomato lentivirus, or PGK-tdTomato lentivirus transduced cells (Figure 1c).

Figure 1. Installation of lentivirus-transduced pig mammary epithelial cells (pMEC) (Rowson-Hodel A R, et al., 2015).

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