A droplet microfluidics based platform for mining metagenomic libraries for natural compounds
Historically, microbes from the environment have been a reliable source for novel bio-active compounds. Cloning and expression of metagenomic DNA in heterologous strains of bacteria has broadened the range of potential compounds accessible. However, such metagenomic libraries have been under-exploit...
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| Format: | Article |
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Multidisciplinary Digital Publishing Institute
2020
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| Online Access: | https://hdl.handle.net/1721.1/125115 |
| _version_ | 1826210497264877568 |
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| author | Theodorou, Elias Scanga, Randall Twardowski, Mariusz Snyder, Michael P. Brouzes, Eric |
| author2 | Massachusetts Institute of Technology. Department of Chemistry |
| author_facet | Massachusetts Institute of Technology. Department of Chemistry Theodorou, Elias Scanga, Randall Twardowski, Mariusz Snyder, Michael P. Brouzes, Eric |
| author_sort | Theodorou, Elias |
| collection | MIT |
| description | Historically, microbes from the environment have been a reliable source for novel bio-active compounds. Cloning and expression of metagenomic DNA in heterologous strains of bacteria has broadened the range of potential compounds accessible. However, such metagenomic libraries have been under-exploited for applications in mammalian cells because of a lack of integrated methods. We present an innovative platform to systematically mine natural resources for pro-apoptotic compounds that relies on the combination of bacterial delivery and droplet microfluidics. Using the violacein operon from C. violaceum as a model, we demonstrate that E. coli modified to be invasive can serve as an efficient delivery vehicle of natural compounds. This approach permits the seamless screening of metagenomic libraries with mammalian cell assays and alleviates the need for laborious extraction of natural compounds. In addition, we leverage the unique properties of droplet microfluidics to amplify bacterial clones and perform clonal screening at high-throughput in place of one-compound-per-well assays in multi-well format. We also use droplet microfluidics to establish a cell aggregate strategy that overcomes the issue of background apoptosis. Altogether, this work forms the foundation of a versatile platform to efficiently mine the metagenome for compounds with therapeutic potential. ©2017 Keywords: metagenomic screening; droplet microfluidics; high-throughput screening; natural compound |
| first_indexed | 2024-09-23T14:50:32Z |
| format | Article |
| id | mit-1721.1/125115 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T14:50:32Z |
| publishDate | 2020 |
| publisher | Multidisciplinary Digital Publishing Institute |
| record_format | dspace |
| spelling | mit-1721.1/1251152022-10-01T22:52:56Z A droplet microfluidics based platform for mining metagenomic libraries for natural compounds Theodorou, Elias Scanga, Randall Twardowski, Mariusz Snyder, Michael P. Brouzes, Eric Massachusetts Institute of Technology. Department of Chemistry Historically, microbes from the environment have been a reliable source for novel bio-active compounds. Cloning and expression of metagenomic DNA in heterologous strains of bacteria has broadened the range of potential compounds accessible. However, such metagenomic libraries have been under-exploited for applications in mammalian cells because of a lack of integrated methods. We present an innovative platform to systematically mine natural resources for pro-apoptotic compounds that relies on the combination of bacterial delivery and droplet microfluidics. Using the violacein operon from C. violaceum as a model, we demonstrate that E. coli modified to be invasive can serve as an efficient delivery vehicle of natural compounds. This approach permits the seamless screening of metagenomic libraries with mammalian cell assays and alleviates the need for laborious extraction of natural compounds. In addition, we leverage the unique properties of droplet microfluidics to amplify bacterial clones and perform clonal screening at high-throughput in place of one-compound-per-well assays in multi-well format. We also use droplet microfluidics to establish a cell aggregate strategy that overcomes the issue of background apoptosis. Altogether, this work forms the foundation of a versatile platform to efficiently mine the metagenome for compounds with therapeutic potential. ©2017 Keywords: metagenomic screening; droplet microfluidics; high-throughput screening; natural compound National Institute of Health (grat no. NCI-1R43GM95227) 2020-05-07T18:31:37Z 2020-05-07T18:31:37Z 2017-07-25 2017-06 2019-03-29T19:40:00Z Article http://purl.org/eprint/type/JournalArticle 2072-666X https://hdl.handle.net/1721.1/125115 Theodorou, Elias, Randall Scanga, Mariusz Twardowski, Michael P. Snyder, and Eric Brouzes, "A droplet microfluidics based platform for mining metagenomic libraries for natural compounds." Micromachines 8, 8 (2017): no. 230 doi 10.3390/mi8080230 ©2017 Author(s) 10.3390/mi8080230 Micromachines Creative Commons Attribution https://creativecommons.org/licenses/by/4.0/ application/pdf Multidisciplinary Digital Publishing Institute Multidisciplinary Digital Publishing Institute |
| spellingShingle | Theodorou, Elias Scanga, Randall Twardowski, Mariusz Snyder, Michael P. Brouzes, Eric A droplet microfluidics based platform for mining metagenomic libraries for natural compounds |
| title | A droplet microfluidics based platform for mining metagenomic libraries for natural compounds |
| title_full | A droplet microfluidics based platform for mining metagenomic libraries for natural compounds |
| title_fullStr | A droplet microfluidics based platform for mining metagenomic libraries for natural compounds |
| title_full_unstemmed | A droplet microfluidics based platform for mining metagenomic libraries for natural compounds |
| title_short | A droplet microfluidics based platform for mining metagenomic libraries for natural compounds |
| title_sort | droplet microfluidics based platform for mining metagenomic libraries for natural compounds |
| url | https://hdl.handle.net/1721.1/125115 |
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