The ultimate goal of our project was to create a toolkit of Site-Specific Recombinases (SSRs) in order to improve their availability and ease of use for other synthetic biologists. We have succeeded in this goal. As demonstrated on our parts page we have assembled Biobrick parts of four tyrosine recombinases (Dre, Vika, VCre, and SCre) and their associated target sites. We have also expanded on the potential of using Cre recombinase biobricks by developing 10 orthogonal Lox mutant sites, which can be recognised by Cre and used to catalyse distinct recombination events in one cell.
We have extensively documented how the parts function, including detailed information about cross-reactivity obtained using our novel measurement constructs. Furthermore, we have obtained novel insight into our recombinase proteins using our measurements constructs, as shown on our results page. Therefore, we can confidently state that our project works: our toolkit has been demonstrated to function, can be used to assemble more complex constructs (for example our measurement devices), and is accompanied by tools like our software designer to make it easy to use. We have also met our goals with our mathematical model. Our model has helped to validate the design of our theoretical constructs and gave useful insight for our collaboration
The toolkit functions, and we were able to demonstrate that it functions under realistic conditions. Our lab-work with the toolkit was in complete compliance with all necessary safety regulations . We have also increased the accessibility of the project, as demonstrated through our integrated human practices. This means that you do not necessarily need to be a recombinase expert to use our toolkit. Therefore, our toolkit functions in a safe manner, and in a way where all synthetic biologists, from high school iGEMers to seasoned P.I.s, can use.