Team:Potsdam/Results
Our results - dCas9
For the dCas9-project we designed a high copy and a low copy plasmid, which should be transformed together in E.coli to do metabolic channelling. We finished our high copy plasmid, so the plasmid with the target sequences for our signal guide RNAs. The plasmid is shown in the figure below:
Our low copy plasmid should include the fusion of the Laci-repressor with dCas9, our enzymes IAAM and IAAH fused with the RNA-binding proteins MS2 and PP7, respectively and the sgRNAs. We finished the pre-plasmids, LacI-dCas9 in pExA2 shown in Figure 2 and IAAM-MS2-IAAH-PP7 (IAA-fusion) in pSB4A5 shown in Figure 3. The next steps to first clone the LacI-dCas9 also in the pSB4A5 with the IAA-fusions and second clone the sgRNA cassettes as last step in this vector, too, we couldn’t finish.
Our low copy plasmid should include the fusion of the Laci-repressor with dCas9, our enzymes IAAM and IAAH fused with the RNA-binding proteins MS2 and PP7, respectively and the sgRNAs. We finished the pre-plasmids, LacI-dCas9 in pExA2 shown in Figure 2 and IAAM-MS2-IAAH-PP7 (IAA-fusion) in pSB4A5 shown in Figure 3. The next steps to first clone the LacI-dCas9 also in the pSB4A5 with the IAA-fusions and second clone the sgRNA cassettes as last step in this vector, too, we couldn’t finish.
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