Synthetic Circuit-Driven Expression of Heterologous Enzymes for Disease Detection
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-assoc...
| Main Authors: | , , , , , , |
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| Other Authors: | |
| Format: | Article |
| Language: | English |
| Published: |
American Chemical Society (ACS)
2022
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| Online Access: | https://hdl.handle.net/1721.1/142861 |
| _version_ | 1826205259854249984 |
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| author | He, Jiang Nissim, Lior Soleimany, Ava P Binder-Nissim, Adina Fleming, Heather E Lu, Timothy K Bhatia, Sangeeta N |
| author2 | Koch Institute for Integrative Cancer Research at MIT |
| author_facet | Koch Institute for Integrative Cancer Research at MIT He, Jiang Nissim, Lior Soleimany, Ava P Binder-Nissim, Adina Fleming, Heather E Lu, Timothy K Bhatia, Sangeeta N |
| author_sort | He, Jiang |
| collection | MIT |
| description | 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. |
| first_indexed | 2024-09-23T13:10:11Z |
| format | Article |
| id | mit-1721.1/142861 |
| institution | Massachusetts Institute of Technology |
| language | English |
| last_indexed | 2024-09-23T13:10:11Z |
| publishDate | 2022 |
| publisher | American Chemical Society (ACS) |
| record_format | dspace |
| spelling | mit-1721.1/1428612024-03-20T18:52:45Z Synthetic Circuit-Driven Expression of Heterologous Enzymes for Disease Detection He, Jiang Nissim, Lior Soleimany, Ava P Binder-Nissim, Adina Fleming, Heather E Lu, Timothy K Bhatia, Sangeeta N 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 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. 2022-06-01T19:48:24Z 2022-06-01T19:48:24Z 2021 2022-06-01T19:41:42Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/142861 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). en 10.1021/ACSSYNBIO.1C00133 ACS Synthetic Biology Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International Licens http://creativecommons.org/licenses/by-nc-nd/4.0/ application/pdf American Chemical Society (ACS) American Chemical Society |
| spellingShingle | He, Jiang Nissim, Lior Soleimany, Ava P Binder-Nissim, Adina Fleming, Heather E Lu, Timothy K Bhatia, Sangeeta N Synthetic Circuit-Driven Expression of Heterologous Enzymes for Disease Detection |
| title | Synthetic Circuit-Driven Expression of Heterologous Enzymes for Disease Detection |
| title_full | Synthetic Circuit-Driven Expression of Heterologous Enzymes for Disease Detection |
| title_fullStr | Synthetic Circuit-Driven Expression of Heterologous Enzymes for Disease Detection |
| title_full_unstemmed | Synthetic Circuit-Driven Expression of Heterologous Enzymes for Disease Detection |
| title_short | Synthetic Circuit-Driven Expression of Heterologous Enzymes for Disease Detection |
| title_sort | synthetic circuit driven expression of heterologous enzymes for disease detection |
| url | https://hdl.handle.net/1721.1/142861 |
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