Our research seeks to apply phage and phage-encoded genes and genetic systems toward the generation of new process innovations with application to the design of novel therapeutics and optimized production/purification platforms.
The bacteriophage PY54 Tel/pal recombinase system has been exploited as a novel platform for the efficient production of linear covalently closed (LCC) DNA minivectors, termed DNA ministrings, to serve as safe and effective DNA delivery vectors. This work aims to construct efficient and scale-able processes to generate DNA vectors in vivo as simplified, scalable and optimized strategies for vector production. In characterizing these DNA vectors, we compared them in safety and effectiveness to conventional DNA delivery vectors. This work has culminated in the development of the most efficient in vivo LCC DNA vector production system to date (Patent awarded US 9290778 B2) and we continue to optimize the efficiency and scalability of the production system (Nafissi et al. 2014. Mol Ther Nucleic Acids 3: e165). See our JoVE video here.
Our lab has designed and characterized a novel 2-dimensional genetic strategy to modulate the degree of decoration of a desired peptide/protein on the surface of phage l via genetic fusion to its major capsid protein, gpD. This was done by combining various permutations of the gpD allele with controllable expression of gpD::X (fusion) during phage assembly. Our studies represent the first method and attempt to measure surface phage fluorescence (eGFP fused to gpD) on a phage by FACS and the highest degree of controllable phage decoration reported to date. (Patent pending US 2015/0031583; Nicastro et al. 2013. Appl Microbiol Biotechnol 97: 7791-7804).