Glioma is the most common primary central nervous system tumor, accounting for about 40-60% of central nervous system tumors. Malignant glioma is one of the chronic diseases of the central nervous system. It has the characteristics of poor prognosis and high recurrence rate. The existing treatment methods such as surgery, radiotherapy and chemotherapy are not satisfactory. In recent years, with the development of molecular biology technology, gene therapy of tumors has become a hot topic in the field of neurosurgery. Zi Yan: "If you want to do something good, you must first sharpen your tools." To study glioma, disease models and genetic manipulation tools are indispensable. So, everyone who wants to “hot spots†has to look at the eyes of Da Nai, and the following content can help you to go shoulder to shoulder with Shenzhou No. 11.
1. Which cell model do I need? Is it easy to operate?
As the saying goes, if the experiment is to be done well, the cells are essential. The cell model has three advantages: a short cycle, a clean background, and easy results, so it is generally the first step in all research. Due to the different characteristics of cells, the difficulty of genetic manipulation is often different. Lentivirus is a very efficient tool for gene manipulation, not only in cell lines, but also in in vivo experiments. So what are the easy-to-operate cell models to choose from in gliomas? The following table summarizes some commonly used cell lines. Each cell is equipped with exclusive experimental data from Jikai.
Second, the cell experiment works well, how does the animal experiment do?
After the cell experiment is done, I want to send a high score article? No confirmation of the in vivo experimental data! In glioma studies, the in vivo model commonly used for proliferation is subcutaneous tumor formation and in situ tumor formation in nude mice.
In addition, a spontaneous glioma model can also be constructed by means of transgenic animals. However, transgenic animals have high cost and long cycle, and their use has great limitations. A powerful introduction to you today is the direct injection of a proto-oncogene lentivirus containing a loxP site into a specific region of the brain of the Cre model mouse, which activates the endogenous signaling pathway and leads to tumor formation.
Third, the in vitro and in vivo models have been selected, then what kind of lentiviral vector does my experiment need to use to manipulate the gene?
Q: How do you take a few steps to get the gene function done?
A: Three steps! Model selection - genetic manipulation - look at the phenotype.
Therefore, after selecting the model, the following genetic manipulation vector needs to be selected. 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. For glioma studies, the GFAP promoter can be used as a tissue-specific promoter. In accordance with the practice, sacrifice a map of God, and immediately customize your exclusive carrier for phenotypic observation. Get high scores SCI, National Natural Science Fund, and embark on the peak of life, just around the corner!
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.
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PS: Friends who want to know more about the Tumor Tools Guide can pay attention to the "Jikai Gene" WeChat public number, and more exciting articles waiting for you.
references
[1] Zhou X, et al. HOTAIR is a therapeutic target in glioblastoma [J]. Oncotarget, 2015, 6: 8353-8365.
[2] Yang Y, et al. Resveratrol Represses Pokemon Expression in Human Glioma Cells [J]. Molecular neurobiology, 2016, 53: 1266-1278.
[3]Xiao DS, etal.Chromatin Remodeling Factor LSH is Upregulated by the LRP6-GSK3β-E2F1 Axis Linking Reversely with Survival in Gliomas. Theranostics, 2017, 7:132-143.
[4] Huang N, et al. Downregulation of nitrogen permease regulator like-2 activates PDK1-AKT1 and contributes to the malignant growth of glioma cells [J]. Molecular carcinogenesis, 2016, 55: 1613-1626.
[5] Yang Y, et al. MiR-129-2 functions as a tumor suppressor in glioma cells by targeting HMGB1 and is down-regulated by DNA methylation [J]. Molecular and cellular biochemistry, 2015, 404: 229-239.
[6] Shir A, Levitzki A. Inhibition of glioma growth by tumor-specific activation of double-stranded RNA-dependent protein kinase PKR [J]. Nature biotechnology, 2002, 20: 895-900.
[7] Zhou H, et al. Dynamic near-infrared optical imaging of 2-deoxyglucose uptake by intracranial glioma of athymic mice [J]. PloS one, 2009, 4: e8051.
[8] Marumoto T, et al .Development of a novel mouse glioma model using lentiviral vectors. Nat Med 2009.15:110–116.
In the previous issues, I have shared a summary of the in vitro and in vivo models and tools of several large cancer species. If you want to see it again, please post the following:
1. Review of in vitro and in vivo models of lung cancer research
2. Review of in vitro and in vivo models of colorectal cancer research
3. Overview of in vivo and in vitro models of gastric cancer research
4. Summary of in vitro and in vivo models of liver cancer
5. Summary of in vitro and in vivo models of pancreatic cancer research
6. Overview of internal and external models of hematological tumor research
7. Breast cancer in vitro and in vivo model summary
8. Review of in vivo and in vitro models of prostate cancer research
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