远红光调控基因表达工程细菌的设计、构建及应用研究

In recent years, the regulation of microbial gene expression and metabolic pathway based on synthetic biology sprang up, the application of which, however, focused on the synthesis and accumulation of natural products. In consideration of the substantial connection between microorganisms and humans, microbial cells have great potential and significance on disease diagnosis and treatment. Therefore, capitalization on synthetic biology and optogenetic approaches, we develop a safe and efficient gene-expression regulation system to achieve high spatiotemporal and precise regulation of gene expression through far-red light, which is then used in light-mediated tumor treatment. First of all, to design and construct the quorum sensing self-driving far-red light-controlled gene expression system, which is optimized and investigated in microbial cells. Secondly, to study the dynamic characteristics of report gene in mice by transplanted the engineered bacteria containing our far-red light-controlled gene expression system. Thirdly, to express cell factors under the regulation of far-red light, and study the efficiency of them on activating the immune system in mice. Finally, the quorum sensing self-driving far-red light-controlled gene expression system will be used for tumor treatment through combinatorial expression of cell factors and anti-tumor proteins, and thus tumor treatment through light will be achieved. This research will provide new strategies and reference for microbial synthetic biology applied on medical research and clinic treatment of tumor.

近年来，以合成生物学为核心调控微生物细胞的基因表达与代谢途径的研究发展迅猛，然而其应用主要集中于天然产物的合成与积累。由于微生物与人体的密切关系，以微生物细胞为载体进行疾病的诊断与治疗具有极大的潜能和意义。因此，本项目结合合成生物学和光遗传学，在微生物细胞中开发了一种安全、高效的光控基因表达调控系统，通过远红光的照射实现时空精准调控基因表达并用于光控肿瘤治疗。首先设计、构建群体感应自驱动的远红光调控基因表达系统，在微生物细胞内进行优化和功能测试；其次，将含有该光控系统的工程菌移植小鼠体内，研究其体内调控报告基因表达的动力学特征；再次，通过远红光调控细胞因子的表达，研究对小鼠免疫系统的激活效率；最后，将该系统应用于肿瘤模型小鼠，通过细胞因子和抗肿瘤蛋白的联合表达协同治疗肿瘤，最终实现光控治疗肿瘤的目的。本项目将为微生物合成生物学在医学研究和临床治疗中的应用提供新的策略与参考。

近年来，以合成生物学为核心调控微生物细胞的基因表达与代谢途径的研究发展迅猛，然而其应用主要集中于天然产物的合成与积累。由于微生物与人体的密切关系，以微生物细胞为载体进行疾病的诊断与治疗具有极大的潜能和意义。因此，本项目结合合成生物学和光遗传学，在微生物细胞中开发了一种安全、高效的光控基因表达调控系统，通过远红光的照射实现时空精准调控基因表达并用于光控肿瘤治疗。首先设计、构建远红光调控的基因表达系统，在微生物细胞内进行优化和功能测试；其次，对底盘沙门氏菌进行减毒改造，提高其用于肿瘤治疗的安全性；再次，将含有该光控系统的工程菌移植小鼠体内，研究其体内调控报告基因表达的动力学特征；最后，通过远红光调控肿瘤杀伤蛋白和纳米抗体的表达，分别应用于冷肿瘤和热肿瘤的肿瘤模型小鼠，实现光控治疗肿瘤的目的。本项目将为微生物合成生物学在医学研究和临床治疗中的应用提供新的策略与参考。