Engineering Synthetic Gene Circuits through Competitive Dimerization.

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Synthetic gene circuits offer considerable potential for engineering human cells to perform complex functions for innovative biotechnological and therapeutic applications. However, current approaches are limited by the lack of versatile, scalable and modular molecular components. DIMERCIRCUITS aims to overcome these barriers by developing a novel class of Modular Artificial Dimerizing transcription factors (MADTFs) whose activity is dynamically regulated through interaction with competitive inhibitory proteins (ΔTFs). These ΔTFs competitively heterodimerize with MAD-TFs preventing their homo-dimerization and thus their function. This dual strategy enables dynamic and reversible regulation of gene expression, establishing a novel technological platform to build synthetic circuits and program cell behaviour. The compact and modular design of MADTFs and ΔTFs not only facilitates their integration into a variety of biotechnological applications but also enhances their potential for clinical translation. Computational modeling and machine learning techniques will be employed to explore the design space of synthetic circuits based on these components, ensuring they meet desired regulatory functions. This framework will guide the experimental implementation of synthetic circuits in human cell lines. As a proof of concept, the therapeutic potential of DIMERCIRCUITS will be demonstrated in patient-derived organoids for the treatment of Friedreich's ataxia, a rare genetic neurodegenerative disorder. By engineering a gene dosage compensation circuit, DIMERCIRCUITS aims to achieve precise regulation of therapeutic gene expression within a narrow window, independent of viral vector uptake, thereby mitigating the overxpression-related toxicity risks associated with current gene therapy approaches. DIMERCIRCUITS aims to establish a foundational technology for precision-engineered synthetic circuits enabling transformative advances in both biotechnology and medicine.