Programmable Synthetic Gene Circuits For The Identification And Suppression Of Hepatocellular Carcinoma Cells

Accurately identifying and killing cancer cells is an important topic in cancer treatment.After years of exploration,scientists have developed an intelligent weapon,called a synthetic gene circuit,that can automatically detect and kill cancer cells.The gene circuit is composed of interrelated genetic switches that not only recognize endogenous cancer biomarkers,but also perform subsequent suicide procedures or express reporter genes in a previously designed manner.Our approach to gene circuits based on post-transcriptional and post-translational regulation is more responsive and demonstrates greater safety than systems based on transcriptional regulation.In this study,we constructed a biosynthetic promoter-free gene circuit with two control layers to improve the specific recognition of tumor cells.We have designed and validated the basic control layer of protein interaction(PPI)sensing using the cleavage of TEV proteases.Another orthogonal control layer is used to detect specific proteins.Two layers of gene circuits are integrated to produce inductive PPIs and specific proteins,forming 10 logic gates.To demonstrate the feasibility of this system,we designed specific gene circuits to detect YAP and 14-3-3σ interactions and alpha-fetoprotein expression in hepatocellular carcinoma(HCC),based on the hepatocellular carcinoma genomic map.If the cells were in AFP expression and HIPPO-OFF state,they would be recognized as HCC cells and release anti-angiogenic single-chain antibodies.Otherwise,the cell will be recognized as a non-HCC cell and will not release effector molecules.Cells carrying the gene circuit distinguish HCC from other cells and release therapeutic antibodies,and the effectiveness of the gene circuit was verified in vitro and in vivo.