<p>Our team will tackle the growing problem of PFAS (poly and per-fluoroalkyl substances) contamination/pollution. PFAS, due to its chemical inertness, are extremely hard to decompose and have been named “forever chemicals”. These substances have become more and more widespread to the point where 97% of Americans have detectable amounts of them in their blood (NHANES 2015). The health risk of PFAS is not fully understood but has been implicated in liver diseases, cancers, increased cholesterol levels, and other complications. Our team attempts to help combat this problem by engineering bacteria to produce an observable signal when they encounter PFAS. PFAS are usually detected with liquid or gas chromatography and mass spectrometry, which is time-consuming, expensive, and inaccessible to the public. PFAS-sensitive bacteria have the potential for extremely high-throughput testing for PFAS. We hope to be able to make such a system based on a genetic circuit transformed into E. coli. We will attempt three different approaches to determining an optimal circuit for PFAS detection. The first uses a previously discovered promoter, prmA, that has been documented to upregulate gene expression in the presence of PFAS. We will test the limit of detection of this promoter to quantitatively measure its efficacy. Second, we will use a gene circuit containing a FAB:GFP conjugated molecule; in the presence of PFAS, this molecule has been shown to change confirmation to allow for the GFP to fluoresce. Finally, we will use a synthetic transcription factor that responds to estradiol in yeast; because PFOA has been shown to be a strong estrogen receptor agonist, we hope that we can recreate the synthetic transcription factor into E. coli. In addition, to better characterize the possible impacts of PFAS, our team is using computer simulations and wet lab testing (via protein assays) to determine different proteins that can bind to PFAS, along with the possibility of computer-aided protein mutations to improve binding. <p>
<p>Our team will tackle the growing problem of PFAS (poly and per-fluoroalkyl substances) contamination/pollution. PFAS, due to its chemical inertness, are extremely hard to decompose and have been named “forever chemicals”. These substances have become more and more widespread to the point where 97% of Americans have detectable amounts of them in their blood (NHANES 2015). The health risk of PFAS is not fully understood but has been implicated in liver diseases, cancers, increased cholesterol levels, and other complications. Our team attempts to help combat this problem by engineering bacteria to produce an observable signal when they encounter PFAS. PFAS are usually detected with liquid or gas chromatography and mass spectrometry, which is time-consuming, expensive, and inaccessible to the public. PFAS-sensitive bacteria have the potential for extremely high-throughput testing for PFAS. We hope to be able to make such a system based on a genetic circuit transformed into E. coli. We will attempt three different approaches to determining an optimal circuit for PFAS detection. The first uses a previously discovered promoter, prmA, that has been documented to upregulate gene expression in the presence of PFAS. We will test the limit of detection of this promoter to quantitatively measure its efficacy. Second, we will use a gene circuit containing a FAB:GFP conjugated molecule; in the presence of PFAS, this molecule has been shown to change confirmation to allow for the GFP to fluoresce. Finally, we will use a synthetic transcription factor that responds to estradiol in yeast; because PFOA has been shown to be a strong estrogen receptor agonist, we hope that we can recreate the synthetic transcription factor into E. coli. In addition, to better characterize the possible impacts of PFAS, our team is using computer simulations and wet lab testing (via protein assays) to determine different proteins that can bind to PFAS, along with the possibility of computer-aided protein mutations to improve binding. </p>
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<h2>References</h2>
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<p>iGEM teams are encouraged to record references you use during the course of your research. They should be posted somewhere on your wiki so that judges and other visitors can see how you thought about your project and what works inspired you.</p>
