A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds
AbstractHistorically, 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. View Full-Text
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Theodorou, E.; Scanga, R.; Twardowski, M.; Snyder, M.P.; Brouzes, E. A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds. Micromachines 2017, 8, 230.
Theodorou E, Scanga R, Twardowski M, Snyder MP, Brouzes E. A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds. Micromachines. 2017; 8(8):230.Chicago/Turabian Style
Theodorou, Elias; Scanga, Randall; Twardowski, Mariusz; Snyder, Michael P.; Brouzes, Eric. 2017. "A Droplet Microfluidics Based Platform for Mining Metagenomic Libraries for Natural Compounds." Micromachines 8, no. 8: 230.
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