Application of lentiviral tools in in vivo and in vitro models in lung cancer research

As the world's largest common tumor, lung cancer is also a hot area of ​​clinical research. According to incomplete statistics, lung cancer-related national natural projects are close to 10% of the total number of cancer projects, and more than 10,000 SCI articles are published each year . Looking at these tenders and articles, the most classic idea is to study the inactivation of a tumor suppressor gene or the activation of proto-oncogenes. Then the question comes, how to effectively operate the target gene according to the needs of the experiment? The ancients said that if the workers want to do their best, they must first sharpen their tools. The understanding of these operating tools can not be underestimated.

Today, I will share with you the application of lentiviral tools in lung cancer research. The 14 years of experience has been accumulated, including the exclusive pre-experiment data of Jikai.

Lentivirus is a tool for achieving efficient cell infection in vitro. Lentivirus can integrate foreign genes into the genome of host cells, and it can stably express with cell proliferation, and has high infection ability to both mitotic and non-dividing cells, combined with its high safety and low toxicity. Lentivirus has become a widely used gene delivery tool in modern cell biology research.

1. Which kind of lentiviral vector is needed for my experiment?

Promoters, fluorescent markers, and resistance tags are the three major factors we need to consider when choosing a carrier. Different components also require corresponding changes in the experiment. The following picture helps everyone sort out the choice of common carriers. If you look at it, is it clear?

Second, what kind of cell model do I need? How is the effect of lentivirus infection?

The tube for lung cancer research can be used without worrying about the unused tool cells. There are various cell lines for adenocarcinoma, squamous cell carcinoma, large/small cells and other lung cancers, and you are no longer afraid of using wrong cells! Each cell is equipped with Jike's exclusive pre-experiment data and references published by the Jikai virus. Come and see it~

*More comprehensive pre-experiment information, please consult the local salesman

Third, the cell experiment works well, how does the animal experiment do?

Animal experiments in lung cancer research are mainly divided into two types: I tumor formation test; II virus direct injection of animals.

I. The operation of the tumor-forming experiment is relatively routine. The virus-infected cells are first used in vitro, and the infected cells are injected into the animal to form tumors. The method of inoculation differs depending on the site of inoculation. The most commonly used lung cancer is subcutaneous and in situ.

*Note: When in situ tumor formation, luciferase is recommended if the surface is not imaged.

II. The virus is injected directly into the animal. Everyone noticed that because of the live injection, the amount of virus that can be injected into animals is limited. Such experiments require higher quality of the virus. The virus used needs to be specially purified to ensure high titer, low toxicity and not easy to cause inflammatory reaction. Here, an intratumoral injection is taken as an example.

Shanghai Jikai Gene Chemical Technology Co., Ltd. was established in 2002 and has a BSL-2 level lentivirus packaging laboratory. The virus production line has passed the ISO9001 quality management system verification, and the monthly average custom genetic virus product packaging has exceeded 1000 times. Lentiviral production uses six QC assays, viral purity fractionation and absolute quantitative detection of viral titers to ensure virus quality.

400 dedicated Jikai people, use professional to change your efficiency.

Fourth, the reference

[1] He D, Wang J, Zhang C, et al. Down-regulation of miR-675-5p contributes to tumor progression and development by targeting pro-tumorigenic GPR55 in non-small cell lung cancer [J]. Molecular cancer, 2015, 14(1): 1.

[2] Li J, Li X, Ren S, et al. miR-200c overexpression is associated with better efficacy of EGFR-TKIs in non-small cell lung cancer patients with EGFR wild-type[J]. Oncotarget, 2014, 5 (17): 7902.

[3] Fu QF, Liu Y, Fan Y, et al. Alpha-enolase promotes cell glycolysis, growth, migration, and invasion in non-small cell lung cancer through FAK-mediated PI3K/AKT pathway[J]. Journal of hematology & oncology, 2015, 8(1): 1.

[4]Chen B, Zhang W, Gao J, et al. Downregulation of ribosomal protein S6 inhibits the growth of non-small cell lung cancer by inducing cell cycle arrest, rather than apoptosis[J]. Cancer letters, 2014, 354( 2): 378-389.

[5] Zeng J, Liu D, Qiu Z, et al. GSK3β overexpression indicates poor prognosis and its inhibition reduces cell proliferation and survival of non-small cell lung cancer cells [J]. PloS one, 2014, 9(3): E91231.

[6] Ni Z, Tao K, Chen G, et al. CLPTM1L is overexpressed in lung cancer and associated with apoptosis [J]. PloS one, 2012, 7(12): e52598.

[7] Zhang S, Liu L, Wang R, et al. MicroRNA-217 promotes angiogenesis of human cytomegalovirus-infected endothelial cells through downregulation of SIRT1 and FOXO3A[J]. PloS one, 2013, 8(12): e83620.

[8] Qi W, Chen J, Cheng X, et al. Targeting the Wnt-Regulatory Protein CTNNBIP1 by microRNA‐214 Enhances the Stemness and Self‐Renewal of Cancer Stem‐Like Cells in Lung Adenocarcinomas[J]. Stem Cells, 2015 , 33(12): 3423-3436.

[9] Wei Shuzhen, Sun Yu, Yang Zhijian, et al. Establishment of orthotopic transplantation model of human EGFP-labeled human lung cancer in nude mice[J]. Chinese Journal of Lung Cancer, 2010 (7): 670-675.

[10]ZHOU Jing, XU Yu, ZHANG Minghui, et al. Inhibition of A549 subcutaneous xenografts by intratumoral injection of lentiviral vector-mediated Bmi1-shRNA[J]. Journal of Third Military Medical University, 2010, 32(20) : 2177-2180.

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