System Overview: The Virus Smart Bomb
Our goal was to make lambda phage specifically target certain E.coli cells—in effect, to make the phage into a “Smart Bomb” that knows which cells to lyse.
Three proteins affect this decision:
1. N antiterminator (lytic vs. nothing)
2. Q antiterminator (lytic vs. lysogenic)
3. Cro repressor (lytic vs. lysogenic)
Our plan was to knock-out the genes for these proteins (one at a time) and add them back under our own regulation with the use of a riboregulator. To do this, there are three subparts of the project which would be stepping stones to achieving this goal:
1. Recombineering into λ-zap: In order to make a riboregulated gene, the wild-type gene must be mutated in the phage and the riboregulated gene must be integrated into the phage genome.
2. Cloning N, Q, and Cro to add them back under our own regulation: In order to regulate the system with each protein, we have to figure out how much N, Q, or Cro is needed to have the E.coli cells switch from mostly lytic to mostly lysogenic.
3. Riboregulator Engineering: The riboregulator will be the actual regulator of the system which determines what ultimately happens to the cells. In order to integrate the riboregulator into the system, an ideal, workable regulator must be engineered into the system so that the appropriate amount of protein (N, Q, or Cro) is produced or repressed.