Difference between revisions of "Team:Northwestern/experiments"

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<h3><center>Packaging Cas9:</h3></center></h3>
 
<h3><center>Packaging Cas9:</h3></center></h3>
  
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<p style="padding-top:2%; padding-right: 15%; padding-left:15%; font-size:14px;" class="big"> <brIn order to successfully package the Cas9 protein into Outer Membrane Vesicles, we needed a source of OMVs.  We transformed our plasmids with saCas9 into a hyper-vesiculating strain of E. coli known as JC8031.  This strain is genetically engineered to create a large amount of OMVs and as a result, any substance whose concentration is high in the periplasm of the cell would also be expected to exist within the generated OMVs. The Cas9 itself also has a His6 tag attached for identification purposes, as well as a signal sequence meant to direct and secrete the protein into the periplasm.</font></p>
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<p style="padding-top:2%; padding-right: 15%; padding-left:15%; font-size:14px;" class="big">In order to successfully package the Cas9 protein into Outer Membrane Vesicles, we needed a source of OMVs.  We transformed our plasmids with saCas9 into a hyper-vesiculating strain of E. coli known as JC8031.  This strain is genetically engineered to create a large amount of OMVs and as a result, any substance whose concentration is high in the periplasm of the cell would also be expected to exist within the generated OMVs. The Cas9 itself also has a His6 tag attached for identification purposes, as well as a signal sequence meant to direct and secrete the protein into the periplasm.</font></p>
  
  

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Packaging Cas9:

In order to successfully package the Cas9 protein into Outer Membrane Vesicles, we needed a source of OMVs. We transformed our plasmids with saCas9 into a hyper-vesiculating strain of E. coli known as JC8031. This strain is genetically engineered to create a large amount of OMVs and as a result, any substance whose concentration is high in the periplasm of the cell would also be expected to exist within the generated OMVs. The Cas9 itself also has a His6 tag attached for identification purposes, as well as a signal sequence meant to direct and secrete the protein into the periplasm.