Difference between revisions of "Team:Northwestern"
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<h2>Our Project</h2> | <h2>Our Project</h2> | ||
| − | <p>Our team is investigating the utility of outer membrane vesicles (OMVs) in the delivery of Cas9 in vivo. All Gram-negative bacteria produce OMVs and use them to deliver toxins, communicate with other bacteria, mediate membrane composition, and extract materials such as metal ions from their environment. OMVs have been successfully engineered to carry heterologous proteins,<a href="#_msocom_1" name="_msoanchor_1" id="_msoanchor_1"><sup>1</sup></a> which makes them an attractive candidate for systems that require the delivery of a functional protein or complex to recipient cells. One such system is the application of CRISPR-Cas9 to treating antibiotic-resistant bacterial infections. At the moment | + | <p>Our team is investigating the utility of outer membrane vesicles (OMVs) in the delivery of Cas9 in vivo. All Gram-negative bacteria produce OMVs and use them to deliver toxins, communicate with other bacteria, mediate membrane composition, and extract materials such as metal ions from their environment. OMVs have been successfully engineered to carry heterologous proteins,<a href="#_msocom_1" name="_msoanchor_1" id="_msoanchor_1"><sup>1</sup></a> which makes them an attractive candidate for systems that require the delivery of a functional protein or complex to recipient cells. One such system is the application of CRISPR-Cas9 to treating antibiotic-resistant bacterial infections. At the moment the most effective methods of delivering Cas9 to target cells are either limited to DNA delivery (bacteriophage delivery<a href="#_msocom_2" name="_msoanchor_2" id="_msoanchor_2"><sup>2</sup></a> and hydrodynamic injection<a href="#_msocom_3" name="_msoanchor_3" id="_msoanchor_3"><sup>3</sup></a>) or require extensive vector engineering in order to facilitate controlled delivery and uptake (lipid-mediated tranfection<a href="#_msocom_4" name="_msoanchor_4" id="_msoanchor_4"><sup>4</sup></a>). We hope to use signal peptides and protein fusions to direct Cas9 protein into OMVs so that the functional complex can be delivered to and act on the resistance genes in antibiotic-resistant bacteria without extensive lipid modification to facilitate uptake.</p> |
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<p><a name="_msocom_1" id="_msocom_1"></a><a href="#_msoanchor_1">[1]</a> Kuehn, Meta J, and Nicole C Kesty. “Bacterial Outer Membrane Vesicles and the Host-Pathogen Interaction.” Genes & Development 19.22 (2005): 2645–55. <a href="http://www.ncbi.nlm.nih.gov/pubmed/14578354" target="_blank">Web</a>.</p> | <p><a name="_msocom_1" id="_msocom_1"></a><a href="#_msoanchor_1">[1]</a> Kuehn, Meta J, and Nicole C Kesty. “Bacterial Outer Membrane Vesicles and the Host-Pathogen Interaction.” Genes & Development 19.22 (2005): 2645–55. <a href="http://www.ncbi.nlm.nih.gov/pubmed/14578354" target="_blank">Web</a>.</p> | ||
| + | <p><a name="_msocom_2" id="_msocom_2"></a><a href="#_msoanchor_2">[2]</a> Yosef I, Kiro R, Molshanski-Mor S, Edgar R, Qimron U. Different approaches for using bacteriophages against antibiotic-resistant bacteria. Bacteriophage. 2014;4(1):e28491. <a href="http://www.tandfonline.com/doi/abs/10.4161/bact.28491#.V3agNJMrIb0">Web</a>.</p> | ||
| + | <p><a name="_msocom_3" id="_msocom_3"></a><a href="#_msoanchor_3">[3]</a> Suda T, Liu D. Hydrodynamic gene delivery: its principles and applications. Mol Ther. 2007;15(12):2063–2069. <a href="http://www.nature.com/mt/journal/v15/n12/full/6300314a.html">Web</a>.</p> | ||
| + | <p><a name="_msocom_4" id="_msocom_4"></a><a href="#_msoanchor_4">[4]</a> Gori JL, Hsu PD, Maeder ML, Shen S, Welstead GG, Bumcrot D. Delivery and Specificity of CRISPR-Cas9 Genome Editing Technologies for Human Gene Therapy. Hum Gene Ther. 2015;26(7):443–451. <a href="http://online.liebertpub.com/doi/10.1089/hum.2015.074">Web</a>.</p> | ||
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Revision as of 16:59, 1 July 2016
Our Project
Our team is investigating the utility of outer membrane vesicles (OMVs) in the delivery of Cas9 in vivo. All Gram-negative bacteria produce OMVs and use them to deliver toxins, communicate with other bacteria, mediate membrane composition, and extract materials such as metal ions from their environment. OMVs have been successfully engineered to carry heterologous proteins,1 which makes them an attractive candidate for systems that require the delivery of a functional protein or complex to recipient cells. One such system is the application of CRISPR-Cas9 to treating antibiotic-resistant bacterial infections. At the moment the most effective methods of delivering Cas9 to target cells are either limited to DNA delivery (bacteriophage delivery2 and hydrodynamic injection3) or require extensive vector engineering in order to facilitate controlled delivery and uptake (lipid-mediated tranfection4). We hope to use signal peptides and protein fusions to direct Cas9 protein into OMVs so that the functional complex can be delivered to and act on the resistance genes in antibiotic-resistant bacteria without extensive lipid modification to facilitate uptake.
References
[1] Kuehn, Meta J, and Nicole C Kesty. “Bacterial Outer Membrane Vesicles and the Host-Pathogen Interaction.” Genes & Development 19.22 (2005): 2645–55. Web.
[2] Yosef I, Kiro R, Molshanski-Mor S, Edgar R, Qimron U. Different approaches for using bacteriophages against antibiotic-resistant bacteria. Bacteriophage. 2014;4(1):e28491. Web.
[3] Suda T, Liu D. Hydrodynamic gene delivery: its principles and applications. Mol Ther. 2007;15(12):2063–2069. Web.
[4] Gori JL, Hsu PD, Maeder ML, Shen S, Welstead GG, Bumcrot D. Delivery and Specificity of CRISPR-Cas9 Genome Editing Technologies for Human Gene Therapy. Hum Gene Ther. 2015;26(7):443–451. Web.
