Designed by: Virginia Massa   Group: iGEM16_WPI_Worcester   (2016-10-14)

Coding region for a constitutively expressed GFP reporter. A section of the GAPDH gene is used as the 5’ UTR, as it is a known successful target site for Cas9.


Figure 1: Wild Type ATG GFP Reporter Fluorescence


Figure 2: Mutated Start Codon GFP Reporter Fluorescence

The data indicates relatively high efficiency in the ATG/ACG reporters. According to the microscopy data the ATG reporter showed 70.57% of fluorescence compared to the positive control, while the ACG reporter only gave 4.21%. Therefore, the mutated start codon is proven to be able to produce the difference between the two versions of the reporter.


Figure 3: Efficiency of ATG/ACG reporters compared to positive control. Error bars represent standard error between normalized results from 3 biological replicates of the experiment.


Figure 4: ATG/ACG reporters efficiency confirmation by flow cytometry. The difference between the wild type and mutant reporter is consistent with the microscopy data.

For how the values were calculated see this:

For the fluorescence values see here: Trial 1 Trial 2 Trial 3