Team:WPI Worcester/Design

The overall goal of our project is to prove that single base nucleotide changes can occur in an organism when editing enzymes are coupled with the CRISPR/dCas9 system. Designing effective plasmids and reporters was the first step in ensuring that we would be able to accurately show this proof of concept.

Reporters

We designed 2 reporters in total, one for each editing enzyme. Each reporter is designed for targeting by the CRISPR/dCas9 complex onto its mRNA due to a 5’ untranslated region. They also both used a green fluorescent protein (GFP) as the editing identifier. When expressed in the cell without presence of the editing complexes, the reporters should not produce GFP. Thus, the cells would not fluoresce. Once the editing complex binds and edits the reporters, however, GFP should then be translated and the cells will glow green.

APOBEC Reporter