• Last year, the SUSTech_Shenzhen team designed a biobrick for cytotherapy. We started our brainstorming in a therapeutic situation. So, we firstly interviewed Dr. Liang Yang from the School of Medicine.
• He introduced the clinical demands about fighting against Pseudomonas Aeruginosa, which was a significant pathogen in nosocomial infections as well as chronic obstructive pulmonary diseases (COPD).
• We then came up with an idea about bacteriophage therapy: It’s promising to modify bacteriophage and biobrick for controlling the invasion of Pseudomonas Aeruginosa, to avoid antibiotic resistance.

• We then started our project design. While searching for the target protein, we found it was interesting that the bacteriophage could develop an anti-CRISPR system to counteract the CRISPR system in bacteria. We started focusing on Aca1 protein expressed in some specific bacteriophages1 which can invade Pseudomonas under the guidance of Dr. Hongda Huang.

Dr. Liang Yang

Professor

Medical Microbiology

School of Medicine, SUSTech

We searched for suggestions about Pseudomonas from different relative fields, including synthetic biology, chemical biology, environmental engineering, and medicine.

• Initially, we tried to inhibit the growth of bacteriophage via virtual-screening molecule to protect Pseudomonas, because Pseudomonasplays a significant role in sewage treatment.
• However,  Shengchun Wu told us that instead of avoiding the invasion of bacteriophage, they utilized bacteriophage to control the dynamic equilibrium of bacteria in the waste-processing system.
• We found that in their industry, they integrated multiple species of bacteria in the activated sludge to construct a system of bioremediation, rather than only using Pseudomonas.
• Based on his suggestions, we looked deeper into biofilm in sewage treatment and understood the complicated components of the microbial community, including Acinetobacter, Aeromonas, Pseudomonas2, and many other microbes. But it’s still possible to find the “Drug” to protect Pseudomonas from excessive phage infection is viable in finding molecules responsible for other microbes in the complex community of biofilm.

Mr. Shengchun Wu

Environmental Engineering

VP

Shenzhen Techand Ecological and Environmental Co. Ltd.

• We consulted Dr. Kai Jiang about chemical genetic approaches to screen our Drug and the possible application of the two-plasmid report system.
• As a chemical biologist, he appreciated this system and he brought up that it was suitable to achieve high throughput screening via 384-well assay plates.
• He also suggested that we can try to utilize certain prepared chemical banks that contain bioactive chemicals with known in vivo targets or even approved drugs. This strategy would certainly be more efficient than screening over a random chemical bank. and we integrated his advice into our further plan.

Dr. Kai Jiang

Research Associate Professor

Chemical Genetics

Academy for Advanced and Interdisciplinary Studies, SUSTech

• We communicated Dr. Yingfei Ma with the anti-CRISPR system and how to optimize the two-plasmid report system. 
• As a synthetic biologist, Dr. Yingfei Ma guided us to analyze different bacteriophages which can be used for testing.
• He was not sure if they got phages code Aca1 protein, but he was willing to look it up.
• He suggested that we might edit an E. coli phage if we were not able to find the phage we wanted.
• Using the edited E. coli phage, we could also “directly” test the efficiency of our drug.

Dr. Yinfei Ma

Professor, Principal Investigator

Synthetic Microbiome

Center of Synthetic Microbiome, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences

• We then interviewed Dr. Liang Yang again. As a specialist in molecular microbiology, he gave us lots of reasonable suggestions about our project：
  • How to optimize our experiment, how to design tests to verify the efficiency of our drug.
• He also talked about two tricky problems with bacteriophage.
  • The first one is that, in the microbiology and structural biology lab, invasion of bacteriophage is common and easily lead to a catastrophe in the pure culture of bacteria. If the drug can be added to the medium, it is able to inhibit the invasion of bacteriophage. It’s similar that, in the biosynthesis and biodegradation industry, like vitamin B12 biosynthesis, such drugs can protect Pseudomonas from the invasion of phage in massive cultivation.
  • For another, he mentioned the application of biofilm in sewage treatment. Commonly, environmental companies prefer to inoculate biofilm into a series of tubes for degrading polluted water. The over-proliferation of biofilm may block the tubes and now the environmental companies can only change for new ones.
• Therefore, if we can utilize the drug to construct a system that can monitor the demand of bacteriophage and then adjust the quantity of biofilm, it may improve the efficiency of environmental companies and help to tackle the environmental problems. We developed a future plan according to his suggestions and tried to discover a proper molecule with a broad-spectrum application based on the existed project.