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   <h3 style="color:white" class="w3-center w3-padding-large"><br><br>Designing 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>
 
<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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             <a class="ch-bubble" href="https://2016.igem.org/Team:Northwestern/Results">LEARN MORE</a></p>
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<p style="padding-bottom:0; padding-right: 10%; padding-left:10%; black; font-size:14px;" class="big"> OMVs are a promising delivery system because of their easy manipulation and broad range of sizes. Our team is working on developing ways to direct the CRISPR system into these export vesicles. To do this, we engineer genetic components that target the Cas9 protein and its guide RNA into a portion of the bacterial membrane called the periplasm. From there, the CRISPR system can be taken up into the OMVs to be delivered to other cells. Upon delivery, the guide RNA will direct the protein to cut out the antibiotic resistance gene. With the gene deleted, the bacteria will not be able to resist antibiotics, making treatments much more effective. NEED TO CHANGE THIS PARAGRAPH + ADD HUMAN PRACTICES BUTTON .<br><br> </p>
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<p style="padding-bottom:0; padding-right: 10%; padding-left:10%; black; font-size:14px;" class="big"> VesiCure aims to create a therapeutic delivery system for CRISPR gene-editing technology using bacterial outer-membrane vesicles (OMVs). We hope to be able to utilize this delivery system to target the antibiotic resistant genes in bacteria, thereby allowing antibiotic treatments to be effective by eliminating the adaptive ability of bacterial resistance. <b>By genetically altering the resistant bacteria, this treatment would help combat the problem of antibiotic resistance directly from the source</b>.<br><br> </p>
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<p style="padding-bottom:0; padding-right: 10%; padding-left:10%; black; font-size:14px;" class="big"> OMVs are a promising delivery system because of their easy manipulation and broad range of sizes. Our team is working on developing ways to direct the CRISPR system into these export vesicles. To do this, we engineer genetic components that target the Cas9 protein and its guide RNA into a portion of the bacterial membrane called the periplasm. From there, the CRISPR system can be taken up into the OMVs to be delivered to other cells. Upon delivery, the guide RNA will direct the protein to cut out the antibiotic resistance gene. With the gene deleted, the bacteria will not be able to resist antibiotics, making treatments much more effective. NEED TO CHANGE THIS PARAGRAPH. </p>
 
  
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Latest revision as of 17:09, 20 November 2017

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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.



Project significance


VesiCure aims to create a therapeutic delivery system for CRISPR gene-editing technology using bacterial outer-membrane vesicles (OMVs). We hope to be able to utilize this delivery system to target the antibiotic resistant genes in bacteria, thereby allowing antibiotic treatments to be effective by eliminating the adaptive ability of bacterial resistance. By genetically altering the resistant bacteria, this treatment would help combat the problem of antibiotic resistance directly from the source.




Check out the video below to learn more about our project!