Synthetic Circuit-Driven Expression of Heterologous Enzymes for Disease Detection
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acssynbio.1c00133.pdf
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Published version
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3.65 MB
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Author(s)
He, Jiang
Nissim, Lior
Soleimany, Ava P
Binder-Nissim, Adina
Fleming, Heather E
Lu, Timothy K
Bhatia, Sangeeta N
Date Issued
2021
Journal
ACS Synthetic Biology
Publisher
American Chemical Society (ACS)
Citation
He, Jiang, Nissim, Lior, Soleimany, Ava P, Binder-Nissim, Adina, Fleming, Heather E et al. 2021. "Synthetic Circuit-Driven Expression of Heterologous Enzymes for Disease Detection." ACS Synthetic Biology, 10 (9).
Version
Final published version
Abstract
The integration of nanotechnology and synthetic biology could lay the framework for new classes of engineered biosensors that produce amplified readouts of disease states. As a proof-of-concept demonstration of this vision, here we present an engineered gene circuit that, in response to cancer-associated transcriptional deregulation, expresses heterologous enzyme biomarkers whose activity can be measured by nanoparticle sensors that generate amplified detection readouts. Specifically, we designed an AND-gate gene circuit that integrates the activity of two ovarian cancer-specific synthetic promoters to drive the expression of a heterologous protein output, secreted Tobacco Etch Virus (TEV) protease, exclusively from within tumor cells. Nanoparticle probes were engineered to carry a TEV-specific peptide substrate in order to measure the activity of the circuit-generated enzyme to yield amplified detection signals measurable in the urine or blood. We applied our integrated sense-and-respond system in a mouse model of disseminated ovarian cancer, where we demonstrated measurement of circuit-specific TEV protease activity both in vivo using exogenously administered nanoparticle sensors and ex vivo using quenched fluorescent probes. We envision that this work will lay the foundation for how synthetic biology and nanotechnology can be meaningfully integrated to achieve next-generation engineered biosensors.
MIT Department
Koch Institute for Integrative Cancer Research at MIT
Harvard University--MIT Division of Health Sciences and Technology
Massachusetts Institute of Technology. Institute for Medical Engineering & Science
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Howard Hughes Medical Institute
Massachusetts Institute of Technology. Research Laboratory of Electronics
Massachusetts Institute of Technology. Department of Biological Engineering
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DOI of Published Version
https://doi.org/10.1021/ACSSYNBIO.1C00133
