Difference between revisions of "Team:East Chapel Hill"

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<p> In many poor countries, excess concentrations of fluoride have entered water sources through erosion of sediment and minerals. These toxic concentrations, which are defined by the World Health Organization to be above 1.0mg/L, may result in an array of health complications. Fluoride has been known to induce cell stress, which in turn impairs the function of ameloblasts who are tasked with forming dental enamel. The resulting disease is dental fluorosis, which manifests in the teeth as porous and yellowed enamel. Other potential consequences of exposure to toxic fluoride levels include skeletal fluorosis, which may result in bone deformities, as well as impaired development. <br>
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Our project involves the development of an operon that will serve as a visual indicator of excess fluoride. We plan to develop and improve the operon created by last year’s iGEM team by experimenting with different fluoride riboswitches and promoters. We hope to find promoters that have an increased gene expression and riboswitches that have increased affinity to fluoride. This would allow for our operon to detect fluoride at levels even lower than 1.0mg/L, a significant improvement from the previous operon. Additionally, we may be able to find different riboswitches that function across a spectrum of fluoride concentrations. By combining them, we could potentially create an indicator that shows the actual amount of fluoride in the water, rather than one that only specifies if the concentrations are above a certain threshold.
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  <h3 style="color:white" class="w3-center w3-padding-large"><br><br>Engineering pathways for integrating functional Cas9 protein into OMVs </h3><br>
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<p style="padding-right: 10%; padding-left:10%; font-size:14px; color:white;" class="big">  Inappropriate use of antibiotics has escalated the growing problem of antibiotic resistance in many threatening diseases. In 2014, the World Health Organization classified antibiotic resistance as a global epidemic. Inactivating resistance genes via Cas9 nuclease-mediated cleavage has been shown to be an effective means of combating this epidemic; however, methods of in vivo delivery are currently limited. Our team aims to deliver Cas9 to antibiotic-resistant, pathogenic bacteria through submicron bacterial outer membrane vesicles (OMVs) as a companion re-sensitization therapeutic to antibiotic treatment. OMVs are naturally produced by all Gram-negative bacteria and are used for crosstalk, stress responses, and nutrient acquisition. Their ability to be modified and directed with relative ease makes them an ideal carrier of CRISPR-Cas9. Aiding conventional antibiotic treatment, our technology will model a complete protein delivery system and transport functional Cas9 to target cells.<br><br>
 
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          <h3 class="ch-bubble">About us</h3>
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          <p class="ch-bubble">Northwestern University iGEM<br>
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          <a class="ch-bubble" href="https://2017.igem.org/Team:Northwestern/Team#">LEARN MORE</a></p>
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          <p class="ch-bubble">Outer Membrane Vesicles<br>
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          <a class="ch-bubble" href="https://2017.igem.org/Team:Northwestern/project">LEARN MORE</a></p>
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          <p class="ch-bubble">Cas9 fusion proteins<br>
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            <a class="ch-bubble" href="https://2017.igem.org/Team:Northwestern/Parts">LEARN MORE</a></p>
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<h3 style="black;" class="w3-center w3-padding-large"><br><br>Our Project</h3><br>
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<p style="padding-bottom:0; padding-right: 10%; padding-left:10%; black; font-size:14px;" class="big">  In many poor countries, excess concentrations of fluoride have entered water sources through erosion of sediment and minerals. These toxic concentrations, which are defined by the World Health Organization to be above 1.0mg/L, may result in an array of health complications. Fluoride has been known to induce cell stress, which in turn impairs the function of ameloblasts who are tasked with forming dental enamel. The resulting disease is dental fluorosis, which manifests in the teeth as porous and yellowed enamel. Other potential consequences of exposure to toxic fluoride levels include skeletal fluorosis, which may result in bone deformities, as well as impaired development. <br>
 +
Our project involves the development of an operon that will serve as a visual indicator of excess fluoride. We plan to develop and improve the operon created by last year’s iGEM team by experimenting with different fluoride riboswitches and promoters. We hope to find promoters that have an increased gene expression and riboswitches that have increased affinity to fluoride. This would allow for our operon to detect fluoride at levels even lower than 1.0mg/L, a significant improvement from the previous operon. Additionally, we may be able to find different riboswitches that function across a spectrum of fluoride concentrations. By combining them, we could potentially create an indicator that shows the actual amount of fluoride in the water, rather than one that only specifies if the concentrations are above a certain threshold.</br> </p>
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  <h3 class="w3-center w3-padding-large">Watch to learn more!</h3><br>
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Revision as of 22:34, 30 June 2018

East Chapel Hill
Home



Engineering pathways for integrating functional Cas9 protein into OMVs


Inappropriate use of antibiotics has escalated the growing problem of antibiotic resistance in many threatening diseases. In 2014, the World Health Organization classified antibiotic resistance as a global epidemic. Inactivating resistance genes via Cas9 nuclease-mediated cleavage has been shown to be an effective means of combating this epidemic; however, methods of in vivo delivery are currently limited. Our team aims to deliver Cas9 to antibiotic-resistant, pathogenic bacteria through submicron bacterial outer membrane vesicles (OMVs) as a companion re-sensitization therapeutic to antibiotic treatment. OMVs are naturally produced by all Gram-negative bacteria and are used for crosstalk, stress responses, and nutrient acquisition. Their ability to be modified and directed with relative ease makes them an ideal carrier of CRISPR-Cas9. Aiding conventional antibiotic treatment, our technology will model a complete protein delivery system and transport functional Cas9 to target cells.



Our Project


In many poor countries, excess concentrations of fluoride have entered water sources through erosion of sediment and minerals. These toxic concentrations, which are defined by the World Health Organization to be above 1.0mg/L, may result in an array of health complications. Fluoride has been known to induce cell stress, which in turn impairs the function of ameloblasts who are tasked with forming dental enamel. The resulting disease is dental fluorosis, which manifests in the teeth as porous and yellowed enamel. Other potential consequences of exposure to toxic fluoride levels include skeletal fluorosis, which may result in bone deformities, as well as impaired development.
Our project involves the development of an operon that will serve as a visual indicator of excess fluoride. We plan to develop and improve the operon created by last year’s iGEM team by experimenting with different fluoride riboswitches and promoters. We hope to find promoters that have an increased gene expression and riboswitches that have increased affinity to fluoride. This would allow for our operon to detect fluoride at levels even lower than 1.0mg/L, a significant improvement from the previous operon. Additionally, we may be able to find different riboswitches that function across a spectrum of fluoride concentrations. By combining them, we could potentially create an indicator that shows the actual amount of fluoride in the water, rather than one that only specifies if the concentrations are above a certain threshold.




Watch to learn more!




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