Part:BBa_K1081000:Experience

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Applications of BBa_K1081000

The expression of dead cas9(dcas9)

figure 1. The construction of the dead cas9(dcas9) included plasmid.




figure 2. The expression of dead cas9(dcas9).
We expressed dcas9 after peomoter P(tetR) BBa_J13002 in pSB1A2 plasmid(figure 1) in E coli strain k12. We use SDS page to show dcas9 was successfully expressed in E.coli(see the figure 2).The molecular weight of dcas9 is 150kDa. The amount of protein of the control(only k12 strain) group euqals to the dcas9 included group. As the arrow indicated, the dcas9 is in the 150kDa spot of the page we run. Combined with different guide-RNAs, we could achieve several stages modulation of the target gene through dcas9-guide RNA complex.

figure 3. Plasmid profile of the partial repression.




figure 4. partial repression schematic diagram

figure 5. The result of partial repression
We use dcas9 combine with a partial repression guide-RNA to block promoter 1 while promoter 2 is still functional. In this experiment promoter 1 is BBa_J23106, promoter 2 is BBa_J23102.

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