A tunable dual-promoter integrator for targeting of cancer cells

Precise discrimination between similar cellular states is essential for autonomous decision‐making scenarios, such as in vivo targeting of diseased cells. Discrimination could be achieved by delivering an effector gene expressed under a highly active context‐specific promoter. Yet, a single‐promoter approach has linear response and offers limited control of specificity and efficacy. Here, we constructed a dual‐promoter integrator, which expresses an effector gene only when the combined activity of two internal input promoters is high. A tunable response provides flexibility in choosing promoter inputs and effector gene output. Experiments using one premalignant and four cancer cell lines, over a wide range of promoter activities, revealed a digital‐like response of input amplification following a sharp activation threshold. The response function is cell dependent with its overall magnitude increasing with degree of malignancy. The tunable digital‐like response provides robustness, acts to remove input noise minimizing false‐positive identification of cell states, and improves targeting precision and efficacy.