A modular synthetic promoter toolkit for programmable gene control in activated and chronically stimulated T cells.
8 affiliations
- 1Center for Advanced Biomaterials for Healthcare, Istituto Italiano di Tecnologia - IIT, Naples, Italy.
- 2Department of Electrical Engineering and Information Technology (DIETI), University of Naples Federico II, Naples, Italy.
- 3Division of Functional Immune Cell Modulation, Leibniz Institute for Immunotherapy, Regensburg, Germany.
- 4Synthetic Biology Center, Department of Biological Engineering, Massachusetts Institute of Technology - MIT, Cambridge, MA, USA.
- 5Medical Clinic and Polyclinic III, Internal Medicine with a focus on Oncology, Hematology and Rheumatology, University Hospital Bonn, Bonn, Germany.
- 6Institute for Bioengineering, School of Engineering, The University of Edinburgh, Edinburgh, UK.
- 7University of Regensburg, Regensburg, Germany.
- 8Center for Advanced Biomaterials for Healthcare, Istituto Italiano di Tecnologia - IIT, Naples, Italy. [email protected].
Abstract
Synthetic circuits that couple transgene expression to endogenous transcription factor (TF) activity enable cellular response to changing functional states. In T cells, antigen stimulation results in dynamic changes of transcription factor expression programs. Harnessing these endogenous TF dynamics for programmable gene control remains an important challenge in engineered T cells. Here, we develop a library of synthetic promoters (SPs) responsive to seven TFs identified through transcriptomic analysis of T-cell activation programs. These promoters couple endogenous TF activity to transcriptional output across a broad dynamic range of inducibility and basal expression. We quantitatively profile promoter performance in HEK293 and Jurkat cells, evaluate them for orthogonality, and integrate them into Boolean logic gate architectures and network topologies to minimize basal transcriptional leakiness and improve ON/OFF ratios. We test selected synthetic promoters in primary human CD8⁺ T cells, under endogenous TF regulation, and we further characterize the NR4A2-responsive promoter subset in vitro and in tumor-infiltrating engineered T cells in a melanoma xenograft model, demonstrating dynamic and reversible activation.Together, these results provide a modular SP toolkit of TF-responsive synthetic promoters for programmable transcriptional control in engineered T cells.
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BibTeX
@article{bonfa2026modular,
title = {{A modular synthetic promoter toolkit for programmable gene control in activated and chronically stimulated T cells.}},
author = {Giuliano Bonfá and Giovanna Martino and Assunta Sellitto and Juan Luis Blazquez-Roman and Fabiana Tedeschi and Antonio Rinaldi and Caio Raony Farina Silveira and Francesca Piro and Fabio Caliendo and Loris Melchiorri and Daniela Perna and Rita Tarnai and Christoph Heuser-Loy and Fabio Mastrogiovanni and Timea Vadász and Freddie Starkey and Evangelos Nikolados and Filippo Menolascina and Luca Gattinoni and Velia Siciliano},
journal = {Nature communications},
year = {2026},
month = sep,
doi = {10.1038/s41467-026-77730-7},
url = {https://europepmc.org/article/MED/42893974},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - A modular synthetic promoter toolkit for programmable gene control in activated and chronically stimulated T cells.
AU - Giuliano Bonfá
AU - Giovanna Martino
AU - Assunta Sellitto
AU - Juan Luis Blazquez-Roman
AU - Fabiana Tedeschi
AU - Antonio Rinaldi
AU - Caio Raony Farina Silveira
AU - Francesca Piro
AU - Fabio Caliendo
AU - Loris Melchiorri
AU - Daniela Perna
AU - Rita Tarnai
AU - Christoph Heuser-Loy
AU - Fabio Mastrogiovanni
AU - Timea Vadász
AU - Freddie Starkey
AU - Evangelos Nikolados
AU - Filippo Menolascina
AU - Luca Gattinoni
AU - Velia Siciliano
PY - 2026
DA - 2026/09/16
JO - Nature communications
DO - 10.1038/s41467-026-77730-7
UR - https://europepmc.org/article/MED/42893974
AB - Synthetic circuits that couple transgene expression to endogenous transcription factor (TF) activity enable cellular response to changing functional states. In T cells, antigen stimulation results in dynamic changes of transcription factor expression programs. Harnessing these endogenous TF dynamics for programmable gene control remains an important challenge in engineered T cells. Here, we develop a library of synthetic promoters (SPs) responsive to seven TFs identified through transcriptomic analysis of T-cell activation programs. These promoters couple endogenous TF activity to transcriptional output across a broad dynamic range of inducibility and basal expression. We quantitatively profile promoter performance in HEK293 and Jurkat cells, evaluate them for orthogonality, and integrate them into Boolean logic gate architectures and network topologies to minimize basal transcriptional leakiness and improve ON/OFF ratios. We test selected synthetic promoters in primary human CD8⁺ T cells, under endogenous TF regulation, and we further characterize the NR4A2-responsive promoter subset in vitro and in tumor-infiltrating engineered T cells in a melanoma xenograft model, demonstrating dynamic and reversible activation.Together, these results provide a modular SP toolkit of TF-responsive synthetic promoters for programmable transcriptional control in engineered T cells.
SN - 2041-1723
ER -