Template:McGill iGEM 2024/Rep Gate Design

The diagnostic results of these modules, which allows the identification of infection-causing bacterial species and associated antimicrobial resistance markers, is reported as a level of fluorescence that increases over time. As such, a general fluorescent reporter for the diagnostic is developed which triggers upon the terminal strand displacement reaction. This reporter part can also be adapted to report the performance of (CasX and SDR amplifier modules) through the use of a DNA translator gate.

This reporter gate consists of a simple 5nt toehold domain with a 15nt branch migration domain. This part is created from two annealed ssDNA strands, Rep[X]-t and Rep[X]-b.

To create the part, a fluorophore-quencher (FQ) pair was covalently linked 5’ and 3’ to the top and bottom oligonucleotides respectively. This linkage is on the 5’ and 3’ non-sticky ends. In McGill iGEM’s design, Rep-t-F, Rep-b-Q are annealed in a 1:1 stoichiometric ratio. The fluorophores used are: