Modular Scalable Synthetic Gene Circuits for Complex Functions Within ‎Minimal Computational Layers in Human Cells

Synthetic gene circuits typically implement sophisticated programs through cascaded ‎computational layers, but these architectures require numerous orthogonal parts, increase genetic ‎payload, and deplete cellular resources, which limits functionality and scalability.‎‏ ‏We developed a modular design ‎framework to engineer scalable gene circuits that execute complex functions within fewer ‎computational layers.

The platform integrates a toolkit to generate orthogonal trans-splicing-based ‎AND gates, native-synthetic hybrid promoters for tunable regulation, and synthetic microRNAs that ‎implement inhibitory logic. Using this approach, we engineered a three-input combinatorial logic gate, ‎a half adder, a full adder, and a dynamic 3-to-1 multiplexer with a dedicated Selector Overload Status ‎output, activated only when both selector inputs are activated.