Overview

During our project, we have thought of innovative ways to deal with Leptospira, as well as raising the awareness of the disease to the general public, providing insights to the industry and useful troubleshooting documents for future researchers. We hope our efforts paid during this project can make a great impact on the scientific community and the development of treating the disease.

Providing insights for future scientific research and development

Novel treatment strategy using miRNA inhibitors instead of antibiotics against Leptospira.

Although antibiotic therapies are available, they often have significant side effects on the human body. Bacterial resistance may also develop after long term usage or incomplete treatment of antibiotics. The miRNA inhibitors can avoid the above issues by the following ways.

Focusing on host defence systems instead of the eradication of bacteria.

Many drugs available nowadays focus on killing the bacteria. In our project, we proposed to boost the host’s immune system to fight against it. This can reduce the harmful effects caused by other chemicals entering our bodies. Aside from therapeutics, the miRNA can also act to prevent Leptospira infection, such a method can also prevent other bacteria which also evades the immune system.

Novel way of boosting the complement system by delivering miRNA through exosomes directly to the organ tissue.

Exosomes are extracellular vesicles generated by all cells that can carry nucleic acids, proteins, lipids, and metabolites. It is the most effective form of implementation due to its great biocompatibility with the body and wouldn’t cause immunological rejection. Furthermore, severe leptospirosis could cause kidney injury and we are proposing using HEK293 kidney cell lines exosomes, which are high affinity on kidney tissue. This would be a new way to enhance the immune system with organ specific-targeted exosomes.

Discovering more potential miRNA biomarkers through database mining.

Research on miRNA biomarkers triggered by Leptospira is very limited. Only a few of the papers include experiments and characterisation. This makes it more difficult in handling our project as well due to the lack of data. Therefore we utilised online databases such as miRDB, miRBase, TargetScan, BLAST and NCBI Gene to find the complementary miRNA with higher target scores on the database of the complement proteins’ 3’UTR with some support of the research papers. Our findings including hsa-mir-144, hsa-mir-4427, hsa-mir-3646, and hsa-mir-630, may be the potential miRNA biomarkers for Leptospira infection and they can narrow the research gap for future science research. The characterised sequences are available on the registry page.

Potential miRNA triggered by leptospirosis that related to the immune/complement system.

We used miRDB, miRBase, TargetScan, BLAST and ncbi to find the complementary miRNA with higher scores of the complement proteins’ 3’UTR with some support of the research papers. We have characterised some of them in our project. We hope that they may become potential biomarkers or treatments for leptospirosis.
pmirGLO_TLR2
pmirGLO_CFH
pmirGLO_CPAMD8

Engaging with the public

Raising awareness of Leptospirosis to the general public

Educating and spreading awareness about leptospira through different IHP activities such as making posters, can allow the public to take more preventive measures to slow the spread of this disease and lower the rate of them being infected. Game stall holding in campus with snacks
Raising awareness during inter-hourse swimming competition Promoting during assembly

Bringing the innovative idea to the industry by getting in touch with professionals

During our project, we have asked many advisors including professors of public hygiene and biochemistry of HKU and CUHK, as well as a gene manufacturing company in mainland China, letting more professionals know about our project and giving them the opportunity to apply our ideas in the industry. Meeting with Professor Xia in CUHK Meeting in HKU

Fostering connection between future Hong Kong iGEM teams

Through several symposiums, and collaborating with other schools in Hong Kong, we can build a closer connection between Hong Kong’s iGEM teams, by understanding each other’s project, as well as promoting the competition.
HKU Symosium
High school symposium in Pui Ching Middle School

Useful dry lab and wet lab documents for future iGEM teams and scientists

A new GROMACS tutorial that includes troubleshooting

During our attempt on using GROMACS for predicting protein interactions in dry lab, we encountered many difficulties and errors.

It has been a struggle for us to understand the codes and get through the bugs. Many of the debug solutions weren’t on the original or the mostly used tutorials which makes it hard for us to continue working. We learned how to debug them by doing much more research online.

Although we decided to not use the software at last due to the complexity of the system and the changes in our project topic, data and it was way beyond our understanding, we made a more detailed tutorial on the basic usage as well as the additional troubleshooting methods. GROMACS is no doubt an advanced software that requires a lot of patience and effort to learn and apply. We hope our tutorial can help other teams to learn this software more efficiently so that they can fasten their progress to utilise their time in other parts of the project as well as spending more time in applying the skill in their project. Hopefully it can help the new iGEM teams at an initial stage, who do not have as much experience, just like us.

We have mastered the basics of how to simulate molecular dynamics in a water environment, this gives us useful resources including:

Energy required for subject to stabilise
Temperature required for subject to stabilise
Pressure required for subject to stabilise
Density required for subject to stabilise
Animation of dynamics of subject in water

Simulate Molecular Dynamics on Different Proteins (in water environment)

A new mini database of models of Leptospira outer membrane protein

Since some of the files needed to run GROMACS are difficult to find on the online tutorials and there’s no protein databases specifically made for Leptospira proteins, we grouped all the files needed together for each Leptospira outer membrane protein. We hope this mini database of Leptospira protein can be accessible to everyone, making it easier to conduct characterisations.

Models of Leptospira outer membrane proteins (Drive)

A new simplified protocol for mammalian cell subculture and transfection for luciferase assay

The protocols we found online were too complex for us to follow. Therefore we have discussed with HKU and together we simplified the protocols for mammalian cell subculture and transfection for luciferase assay. We hope this protocol can help future iGEM teams in handling these experiments in a more easier way.

Protocol for mammalian cell subculture and transfection for luciferase

Link to Parts