How we performed our human practice
How the advice from experts changed our projects
Our goal for this year is to develop a biosensor that quantifies HHV-6, and we wanted to propose that the quantification of HHV-6 by our device be incorporated into health checkups. By using this device, people undergoing medical checkups can recognize not only their physical health condition but also their level of fatigue as an objective, thereby preventing mental and physical illnesses caused by fatigue. Our ultimate goal was to make such a device easy to use by people without biological knowledge or training in experimentation. We first identified the Toehold switch, Aptamer, and Cas proteins as potential biosensors for quantification, and created a development workflow for each to discuss which biosensors would be most suitable for our project and more feasible for social implementation. We discussed which biosensor is most suitable for our project and which is more feasible for social implementation.
Getting suggestions from Professors
Meeting with Prof.Gu
On February 19, 2020, we interviewed Professor Man Bock Gu of Korea University, who was temporarily visiting Gifu University as a visiting professor. Professor Gu is a specialist on aptamer, who is the first person in the world to successfully screen aptamers without immobilizing the target using graphene, a new nanomaterial, as a method for selecting aptamers. During our discussion with Prof. Gu, he suggested that detection using a pair of aptamers might be suitable for our project. A pair of aptamer-based detection has the advantage of using a secondary signal aptamer to generate an easily and highly amplified signal. This sandwich binding method was limited in its ability to fully adapt to all target molecules due to the inability to screen the available a pair of aptamers. This is because in the usual SELEX process, the target is fixed on a solid support, which limits the exposure of the target surface and makes it very difficult to separate the two aptamers that bind to different sites on the target simultaneously. However, Prof. Gu's group developed a new SELEX process using graphene oxide that can obtain a pair of aptamers by an immobilization-free of target method, and since then, many SELEXs have been developed by improving this method.
Getting advice from Dr. Nagai and Prof. Shichiri
We first listed Toehold switch, Aptamer, and Cas protein as biosensors to perform the quantification of HHV-6 for our project this year. Then we created a workflow for each of them to find out which biosensor is most suitable for our project. As one of the project proposals, we considered quantifying HHV-6 by Toehold switch using microfluidics. We predicted that this method has advantages compared with real-time qPCR in performing amplification of target DNA, transcription, and translation isothermally and it can detect target DNA more quickly. In order to examine this proposal, we discussed Prof. Hidenori Nagai and Prof. Motoki Shichiri who are in the National Institute of Advanced Industrial Science and Technology (AIST), experts in stress markers and stress quantification. Prof. Nagai is a specialist in micro/nano-device research and has been involved in the development of microfluidic systems for stress measurement. Prof. Shichiri previously worked in the Stress Signaling Group of the Health Engineering Research Division at AIST, where he has explored biomarkers useful for evaluating mental stress and early diagnosis of various diseases.We discussed whether it is possible to perform the reaction on microfluidics and how we could construct a more superior system to real-time qPCR. In addition, We consulted whether HHV-6 is suitable as a biomarker in our project. Prof. Shichiri advised us that HHV-6 would be an appropriate target for this project since he has detected an increase in the amount of HHV-6 based on fatigue. The discussion also revealed that microfluidics do not match our project for the following reasons
For these reasons, we dropped the idea of using microfluidics to quantify HHV-6. Thus, we considered a more simple method to quantify HHV-6 by Cas Protein as a next scheme. Following the advice and suggestions, we decided to apply the dsDNA detection method called DETECTR developed by doudna in 2018.
Learning the current situation from Prof. Horita
We visited Prof. Ryo Horita, Assistant Professor of the Department of Psychiatry and Neurology at Gifu University and a clinical psychologist, to discuss the effectiveness of our device, and its use in health checkups. Prof. Horita works as a clinical psychologist at the Gifu University Health Service Center, where he provides mental health care to patients by counseling them to find out the cause of their stress and giving them advice on how to solve it. According to Prof. Horita, since clinical psychologists and psychiatrists value face-to-face consultations with patients, there has been no approach to understand the mental health of patients by quantifying fatigue. However, he also said that quantifying the mental health status and visualizing the fatigue is very useful to make the medical treatment smoother and also to make patients aware of their own fatigue. In particular, there are many typical Japanese people who are overwhelmed with work and do not acknowledge their fatigue even though it is accumulating. The device we've designed can give such people an objective indicator of their level of fatigue. He also said that one of the most important aspects of using such a system in the field is its immediacy. He suggested that if the results can be shown to patients who come for counseling on the spot and can be linked to the next treatment, the system will be widely used in the field. Dr. Hotta appreciated that our device could be used in the real world in this respect since we could get results in a relatively short time. In addition, he mentioned that such a device has not been used so far because the following two reasons;
Our biosensor can be used with less prior knowledge than other biosensors since it is based on DETECTR. In addition, the detection is very simple to operate. We believe that our device overcomes the second problem. However, He indicates that it would be very hard to use our device in a standard for health checkups and medical diagnosis since it is necessary to obtain permission from the Ministry of Health, Labor and Welfare in Japan if it is used as medical treatment. It requires an enormous amount of time and funds. For this reason, we got to know that our final goal was very difficult to achieve. Discussions with Prof. Horita revealed that the design of the DETECTR-based device is suitable for use in the field, since it can be operated easier and quicker than other biosensors. On the other hand, it became clear that it would be difficult to use for medical purposes such as health checkups due to legal restrictions. Therefore, we have come to the conclusion that the final application of our system should not be in the medical field, but rather change the way it is operated to a device whose purpose is to maintain health in daily life. We also received a suggestion that if this project is successful, it could be used as a project for a health promotion seminar to be held at Gifu University.
Interacts with public
We had a seminar with Kinjirushi Corporation on November 2, 2020. Kinjirushi is a company that produces and sells wasabi. This company is one of the most famous wasabi companies in Japan, holding a 40% share of the commercial wasabi market in our country. Recently, Kinjirushi has been working to promote the appeal of Wasabi to the world and is interested in global initiatives. Through this seminar, we learned about the effects and characteristics of Wasabi, which we did not imagine. On the other hand, we explained about synthetic biology, the competition of iGEM, and the research project of our team to the member of Kinjirushi. The member from Kinjirushi had never known synthetic biology before this seminar, though through this exchange, they were able to notice the potential of synthetic biology and became interested in iGEM. From this activity to now, Kinjirushi Co. supports ourproject. We believe that we were able to increase the number of people who are attracted to iGEM and synthetic biology and want to develop them further.
Open Campus of Gifu University
We released an introductory video about iGEMGifu on Gifu University's YouTube channel as an online open campus of Gifu University on June 10, 2021. Open Campus is an event that Universities in Japan invite high school students to introduce school life. In addition to the activities and attractions of iGEM, each member of our team talked about why we joined iGEM. We believe that by actively communicating using such a major tool, we were able to increase opportunities for people of all ages to learn about iGEM and boost the synthetic biology community. We also believe that this project has appealed to students of Gifu University and also to high school students who are considering entering Gifu University. We hope that people who watched this video will take an interest in biology and our activities through this exhibition. The short version of our video is as below.