Kashkin Kirill N, Chernov Igor P, Didych Dmitry A, Sverdlov Eugene D
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow, Russia.
Biotechniques. 2017 Sep 1;63(3):107-116. doi: 10.2144/000114586.
Gene therapy is a fast-developing field of molecular medicine. New, effective, and cancer-specific promoters are in high demand by researchers seeking to treat cancer through expression of therapeutic genes. Here, we created a combinatorial library of tumor-specific chimeric promoter modules for identifying new promoters with desired functions. The library was constructed by randomly combining promoter fragments from eight human genes involved in cell proliferation control. The pool of chimeric promoters was inserted into a lentiviral expression vector upstream of the CopGFP reporter gene, transduced into A431 cells, and enriched for active promoters by cell sorting. The enriched library contained a remarkably high proportion of active and tumor-specific promoters. This approach to generating combinatorial libraries of chimeric promoters may serve as a useful tool for selecting highly specific and effective promoters for cancer research and gene therapy.
基因治疗是分子医学中一个快速发展的领域。寻求通过治疗性基因表达来治疗癌症的研究人员对新型、有效且具有癌症特异性的启动子有很高的需求。在此,我们创建了一个肿瘤特异性嵌合启动子模块组合文库,用于鉴定具有所需功能的新启动子。该文库通过随机组合参与细胞增殖控制的八个人类基因的启动子片段构建而成。将嵌合启动子库插入到CopGFP报告基因上游的慢病毒表达载体中,转导至A431细胞,并通过细胞分选富集活性启动子。富集后的文库包含比例极高的活性和肿瘤特异性启动子。这种生成嵌合启动子组合文库的方法可能成为为癌症研究和基因治疗选择高度特异性和有效启动子的有用工具。