<p> The funding provided by the IPRD will be utilized for the next 6 months after the iGEM Jamboree, testing the hypothesis that: <b>"A TMA Oxidation Cell converts TMA to TMAO at therapeutic levels for TMAU patients."</b> The specific aim of the proposed project is to rigorously test the performance of the TMA Oxidation Cells in an in vitro model of the human small and large intestines. Below are the steps to be continued after the Jamboree.</p>
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<li><b>Step 1 (1 Month)</b> – Further Develop Proof-of-Concept by building new TMA Oxidation Cells A, B, and C using E. coli Nissle as the host cell that is an FDA-approved probiotic strain.</li>
<li><b>Step 2 (3 Months)</b> – Develop a TMA Oxidation Assay to measure the levels and rates of TMA oxidation by TMA Oxidation Cells A, B, and C in simulated gastrointestinal fluid and ambient oxygen tension using High Performance Liquid Chromatography (HPLC) [20]. Measure the intracellular levels of the Roseovarius sp 217 TMM, R. pomeroyi TMM, and Human FMO3 using the Agilent Bioanalyzer 2100, a Microfluidic Gel Electrophoresis System in the FSU College of Medicine Translational Science Laboratory </li>
<li><b>Step 3 (2 Months)</b> – Extend the TMA Oxidation Assay to measure the levels and rates of TMA oxidation by TMA Oxidation Cells A, B, and C in simulated gastrointestinal fluid and low oxygen tension. Select the best performing cell for further improvements. </li>
<li><b>Expected Outcome</b> - The expected impact of accomplishing the aim is the delivery of an engineered cell that is a candidate cell-based therapeutic for TMAU that could proceed towards in vivo testing. </li>
