Team:Carnegie Mellon/Project/Future

Future Directions

This project laid down the framework for producing temperate phages capable of killing bacteria, thereby circumventing the issue of lysogeny for phage therapies. However, KillerRed as it exists in the Parts Registry is not codon optimized for bacteria and therefore it may be difficult to get robust expression from a single or low copy gene (in the case of a lysogen). We have been able to demonstrate expression of mRFP1 in the prophage but due to the nature of the KillerRed protein, similar functional protein levels were not obtained. Stronger promoters and ribosome binding sites can be utilized for higher expression of a codon-optimized, monomeric¹, KillerRed gene in a prophage to test for killing and therefore, potential therapeutic phage applications. Different temperate phages can be engineered as well to target different organisms so that a cocktail of phages can be produced to kill pathogenic bacteria.

References

¹Takemoto K, Matsuda T, Sakai N, Fu D, Noda M, Uchiyama S, Kotera I, Arai Y, Horiuchi M, Fukui K, Ayabe T, Inagaki F, Suzuki H, Nagai T. 2013. SuperNova, a monomeric photosensitizing fluorescent protein for chromophore-assisted light inactivation. Sci Rep. 17(3):2629.