Programmable Synthetic Protein Circuits for the Identification and Suppression of Hepatocellular Carcinoma

Abstract Precisely identifying and killing tumor cells are diligent pursuits in oncotherapy. Synthesized gene circuits have emerged as an intelligent weapon to solve these problems. Gene circuits based on post-transcriptional regulation enable a faster response than systems based on transcriptional regulation which requires transcription and translation, showing superior safety. In this study, Synthetic-promoter-free gene circuits possessing two control layers were constructed to improve the specific recognition of tumor cells. Using split-TEV, we designed and verified the basic control layer of PPI sensing. Another orthogonal control layer was built to sense specific proteins. Two layers were integrated to generate gene circuits sensing both PPI and specific proteins, forming ten logic gates. To demonstrate the utility of this system, the circuit was engineered to sense AFP expression and the PPI between YAP and 14-3-3σ, the matching profile of HCC. Gene circuit loaded cells distinguished HCC from other cells and released therapeutic antibodies, exhibiting in vitro and in vivo therapeutic effects.