National Research Center "Kurchatov Institute", 123182 Moscow, Russia.
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, 117997 Moscow, Russia.
Int J Mol Sci. 2023 Apr 13;24(8):7192. doi: 10.3390/ijms24087192.
The identification of tissue-specific promoters for gene therapeutic constructs is one of the aims of complex tumor therapy. The genes encoding the fibroblast activation protein () and the connective tissue growth factor () can function in tumor-associated stromal cells but are practically inactive in normal adult cells. Accordingly, the promoters of these genes can be used to develop vectors targeted to the tumor microenvironment. However, the efficiency of these promoters within genetic constructs remains underexplored, particularly, at the organism level. Here, we used the model of embryos to study the efficiency of transient expression of marker genes under the control of promoters of the , , and immediate early genes of (CMV). Within 96 h after the injection of vectors, the and CMV promoters provided similar equal efficiency of reporter protein accumulation. In the case of the promoter, a high level of reporter protein accumulation was observed only in certain zebrafish individuals that were considered developmentally abnormal. Disturbed embryogenesis was the factor of changes in the exogenous promoter function. The data obtained make a significant contribution to understanding the function of the human and promoters within vectors to assess their potential in gene therapy.
鉴定组织特异性启动子用于基因治疗构建是复杂肿瘤治疗的目标之一。成纤维细胞激活蛋白(FAP)和结缔组织生长因子(CTGF)的编码基因可以在肿瘤相关的基质细胞中发挥作用,但在正常成年细胞中实际上是不活跃的。因此,可以利用这些基因的启动子来开发针对肿瘤微环境的载体。然而,这些启动子在遗传构建体中的效率仍未得到充分探索,特别是在机体水平上。在这里,我们使用 胚胎模型研究了标记基因在 FAP、CTGF 和即刻早期基因 (CMV)启动子控制下的瞬时表达效率。在注射载体后的 96 小时内, 和 CMV 启动子提供了相似的报告蛋白积累效率。在 启动子的情况下,仅在某些被认为发育异常的斑马鱼个体中观察到报告蛋白积累的高水平。胚胎发生紊乱是改变外源 启动子功能的因素。获得的数据为理解人 和 启动子在载体中的功能做出了重要贡献,以评估它们在基因治疗中的潜力。