Our Solution
Engineering a phage to increase its host range is one of the fundamental barriers, that is focused on increasing the range of bacteria it can infect.
BRED, or Bacteriophage Recombineering on Electroplated DNA, is a highly effective homologous recombineering method that exploits the natural mechanism of phages: mutations. It works by inserting DNA into specific points of the host phage's genome to cause mutations to the binding receptors. Now we would test this phage's efficacy of lysing the targeted bacteria.
We can collet the plaque from the lysis of the bacteria, and using Polymerase chain reaction, we can purify and analyze it to isolate the mutation. The successful mutation is analyzed against a naturally occuring mutation through polymerase chain reactions (PCR). The main goal is to identify exactly where the point mutations occured, how effective they were and their easibility for extraction.
All of this can ultimately revolutionize the process of discovering the most effective and viable mutation to increase the range of the phage.
Bacteriophages can be used as a novel and efficient category of gene delivery vehicles for the introduction of various diagnostic and therapeutic cargoes to human cells. Using phages as delivery mechanisms are not toxic and pose no side effects.
These phages inject their genetic material into bacteria by recognizing the lipopolysaccharides, pili, peptidoglycans, proteins and teichoic acids comprising the cell walls and outer membranes of the bacteria.
In our solution, transducing particles are used as an antimicrobial agent that can go beyond lytic phage therapy. Particles are primarily used to transfer the inhibited resistance mechansism genes to other strains of bacteria. The assembly of transduction particles is comprised of the nucleotide cargo (inhibited resistance mechansisms), packages inside a viral protein.
Once a phage would infect a bacteria, it can use the bacterial chromosome to produce transduction particles to make the bacteria’s resistance mechanisms weaker, assist in the bacteriophage lysis process and code antigen production instructions for the dendritic cells. Upon lysis, the transduction particles would be realeased and continue the process, leaving us with weakened bacterial ressitance mechanisms and dramatically reduced growth rates in the majority of the remaiing pathogenic bacteria.
Traditionally, when a bacteriophage attacks a bacteria, and innate immune response is triggered and phagocytes are sent to kill the pathogenic bacteria. While this is effective in the short term, there is no memory of the bacteria.
This is the job of the adaptive immune system, which focuses on the development of antibodies against pathogens through identification of unique antibodies. Our solution uses the transduction particles mentioned in the previous step to activate the production of antibodies against the bacteria. Triggering an adaptive immune response is the main alternative to antibiotics, the main method of killing the bacteria.
The first step is to use phage display technology to identify required antigens for the bacteria we are targeting through mapping phage-peptide relationships. Through a process called biopanning, the correct binding antigen is selected, washed and checked to see whether it can be used with the phage. After that, would engineer those specific antigens on the surface of the transduction particles.
The key to activating the adaptive immune system for a bacterial infection is the bacteriophages. Phages naturally have an affinity towards B lymphocytes, which will attract them to the location. Once the lymphocytes engulf the transduction particles, they will identify the antigens and activate the production of antibodies against those antigens, and therefore against the bacteria.
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