Synthetic Circuit-Driven Expression of Heterologous Enzymes for Disease Detection
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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
Journal
ACS Synthetic BiologyTL;DR
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Funding
Howard Hughes Medical Institute FundingLeadership, Planning and EvaluationCommunity Outreach Education and Engagement CoreMolecular Biophysics Training GrantCommunity Outreach Education and Engagement CoreHHS | NIH | National Cancer Institute Grant P30-CA14051Virginia and D.K. Ludwig Fund for Cancer Research FundingMolecular Biophysics Training GrantNational Science Foundation FundingAmar G. Bose Research Grant FundingKoch Institute Marble Center for Cancer Nanomedicine Funding