Team:Northwestern/experiments
Packaging Cas9
In order to successfully package Cas9 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.
