At the beginning of our iGEM activities, we felt that people seem to be unfamiliar with synthetic biology and its social contribution. In order to solve these issues, we chose to show our concepts through Snow White, one of the most famous stories around the world. Besides, we held many outreach activities, dialogued with experts, and created a model called "3E": education, ethics, and economy. We also designed some ideas based to our research (Toxin-Antitoxin system etc.) for society.
We contributed to raising public awareness of synthetic biology and improving our project through science communication.

We approach the social meaning of our project from three aspects: Education, Ethics, and Economy. Education plays an important role to raise scientific literacy as one of the most fundamental activities in daily life. Ethics is indispensable when you are engaged in synthetic biology. Economic profit is produced by technological application of synthetic biology. To approach the social meaning (of our project) from these aspects, we worked on many activities. This page shows all of our human practice activities.

Overview

We consider the possibility of the protein production system is controlled by Maz system and signal transfer mechanism. To improve the performance of controlling the production by Maz system, we developed a tool for increasing or decreasing the number of ACA sequence, which is the specific site cleaved by MazF homo dimmer. Maz system.

1. Controlling the protein production

2. Selectivity of the target mRNA

3. System of communicating with outside

Combining i. and ii., it is expected that the target protein is obtained effectively and with favorite rate. Furthermore, adding iii., we could design an automatic and advanced information-processing system. In other words, we could design an oscillatory cell-cell communication in which one cell containing man system, receives an extrinsic signal (Maz system is controlled by the type of the signal), then the cell produces the substance which affects other cells.

To increase the selectivity of repressing the protein production by Maz system, we develop a tool of increasing or decreasing the number of ACA sequence named "ACA Dwarfs." This application makes our future work much more realistic than other iGEM teams (see Model page).

Furthermore, a lot of vital phenomena are oscillatory. So we could apply this oscillatory system to DDS (Drug Delivery System) or to decrease the risk of the appearance of the multi-drug-resistant strains.

• Decrease the risk of the appearance the multi-drug-resistant strains

  These days, the possible appearance of multi-drug-resistant strain due to excessive use of pesticides is alarming. But spreading a pesticide one by one costs money, so we suggest using the system of controlling the protein production a by Maz system: the production of a pesticide is usually repressed by MazF, but only when the substance delivered from the targeted organism (for example: pheromone), MazF is contradicted by MazE and the pesticide is produced. This oscillatory model can decrease the possibility of the appearance of the multidrug-resistant strains because the pesticide is produced at intervals.

• DDS(Drug Delivery System)

  Introducing maz system to human cells and orchestrating the human original signal transduction system and Maz system, we design the mechanism that the cell secrets drug depended on the internal condition. For example, when a maz system introduced cell receives the blood glucose level increase, the cell synthesizes the insulin.

• Our TA systems and QS might be applicable to control the Intestinal Flora

  1.Introduction

  These days, Intestinal Flora draws a lot of attention because it will greatly affect our health. If the balance of intestinal flora is lost, it will lead into an illness or might even change your personality. It is assumed that intestinal flora consists of communication systems among various bacteria. However, no such communication systems have been found. So, it will contribute to the society to clarify the mechanism of intestinal flora and to find a way of controlling it.

  Our project focused on the cell-cell communication through TA system and QS, and proved the initial step to explain and control the system of the intestinal symbiosis.

  

  2. The results of our project

  2-1. TA System Assay

  We used Maz system, which is a kind of TA system. MazF (Toxin) inhibits translation by cleaving ACA sequence in mRNA and MazE (Antitoxin) releases MazF inhibition. We showed that Maz system could regulate cell growth when mazE and mazF were alternately expressed in E. coli (see TA system assay pages).

  
  

  2-2. AHL and AmiE Assay

  We indicated that QS of Pseudomonas aeruginosa genesand V. fischeri, and amiE, which selectively degrades AHLs, could work in E. coli. Sometimes, these genes are toxic to E. coli, but this was solved by, for example, tagging ssrA or experimenting at low temperatures (see AHL reporter assay page and AmiE assay page).

  
  

  2-3. Temperature-dependent Promoters

  We indicated that temperature-dependent promoters were activated strongly at a specific temperature: gfp under Pcold, which is activated at 15°C and under, was much more expressed at 18°C than at 37°C; on the other hand, gfp under Pheat, which is activated at 42°C and over, was more expressed at 37°C than at 28°C. We can say that we can control the expression of the gene under these temperature-dependent promoters by changing temperature.

  
  

  2-4. Modeling

  We simulated our final genetic circuits containing the genes of TA system, QS and temperature-dependent promoters. The simulation showed that the circuits could perform favorably by improving some parts (see Model page and Rhl system assay page).

  3. Discussion

  In conclusion, we proved that the genes of other bacterial QS and temperature-dependent promoters worked in E. coli and that E. coli original TA system gene under inducible promoters regulated the growth in WET experiments. From these results, the simulation showed that E. coli containing TA system gene and QS gene under various inducible promoters communicated with each other.
  Besides, by simulating that the genetic circuits worked properly with the three types of inducible promoters, we suggest that intestinal bacterial growth is controlled by nutritional condition, temperature and intestinal bacterial density. The three types of promoters were the sugar-dependent promoters (PBAD, Plac), the temperature-dependent promoters (Pcold, Pheat) and the AHL-dependent promoters (Prhl, Plux and Plas). Our project therefore helps to understand the symbiosis of intestinal flora. Furthermore, we are quite sure that we could engineer the E. coli or other bacteria which have function of regulating the symbiosis.

During promotion of our project, we had dialogues with the public and experts. Based on the opinions from them, we developed our project. Keeping from our narrow view, this led to success in creation of a well-rounded project with connection between the public and experts.

This page explains changes that some events had made to our project in chronological order.

Human Practices

Integrated Human

Our Project

Practices

Public Engagement / Dialogue
May Festival

At the university festival of University of Tokyo, the public (parents and friends of students at University of Tokyo, and the other visitors of the festival) advised on our project in a poster session.

Among the projects, the ones with approachable themes were quite popular.

Select "Snow White" from prospective projects

Snow White is the story everyone knows, and people can understand the story pretty much. Moreover, it is familiar to the public and promotes a better understanding about E. coli, synthetic biology and our project.

Public Engagement / Dialogue
Suzukake Festival

At the university festival of Tokyo Tech, the public (parents and friends of students at Tokyo Tech, and the other visitors of the festival) advised on our project in a poster session. We got an advice that we should put on a shortplay to make our story easier to be understood.

Let's perform a short play during the presentation!

Public Engagement / Dialogue
Homecoming Day

This is an annual event held at Tokyo Tech with everyone related to Tokyo Tech and neighboring residents, where we aimed to get feedback from them by introducing our activities with a poster. However, we could hardly tell the public science and technology easily, and they had trouble understanding fundamental knowledge.

• Make videos about E. coli and synthetic biology for the public.
• Make an interesting card game for study.

School-visits
Ochanomizu University Junior High School

Aim of the lesson was to obtain feedback on our project from junior high school students, and to get evaluations of our project on the basis of their criteria. One student offered a following comment.
"Don't other characters appear in the story?"

At the beginning, the characters were three: Snow White, the Queen and the Prince. To make our story closer to the original one, we added the Magic Mirror to the story.

There are some versions of Snow White story. Using our parts, we designed two gene circuits to represent different story versions.

School-visits
Tokyo Metropolitan Nishi High School

Aim of the lesson was to raise student’s interest in iGEM through explanation of possibilities of genetic modifications, gene recombination techniques, and synthetic biology. The other aim of this lesson was to get feedback on our project from high school students. In the lesson, we focused on explanation of our project at that time. One student gave the following opinion.

"Certainly, representing Snow White Story is interesting, but can you make your project work for society?"

Can we make something socially beneficial?

We discuss by ourselves, and set up opportunities to have dialogues with experts.

Expert Prof. Nakasaki

During the dialogue, he said that repetition of protein production control probably takes a step to the technology reducing risk of the multiple-drug-resistant bacteria emergence.

Open

We got the positive opinion that TA system probably leads to a valid technology in the future.

However, there are two problems with protein production control using TA system.
(i), You cannot produce target protein selectively; you produce other needless protein simultaneously.
(ii), Repetition of protein production control leads to sustainable production in one system. However, to take protein from E. coli, you have to destroy E. coli, which makes sustainable production impossible at present.

We continued the activities to find the clue of two critical problems. We worked on them (i) from a view of modeling, (ii) from a view of experiments, and finally found solutions for the problems.

1. "ACADwarfs"
   The development of the software which adjusts the number of ACA (Adenine-Cytosine-Adenine) sequence in the DNA.

   This makes it possible to adjust the inhibition of each protein, that is, you can produce a bigger ration of target protein when MazF works. Moreover, we designed the Java code so that many people can use it.

2. The experiments to make E. coli mutants that secrete protein extracellularly and "TA system ~the Queen's Caprice~"

   UV irradiation has being performed to make the mutants that secrete protein extracellularly.

Using "ACADwarfs" gets the higher grade target protein, which realizes high performance material production system. Additionally, preventing mRNA cleavage of target protein and cleaving mRNA of unnecessary protein can establish sensitive protein labeling.

Furthermore, our research leads to DDS (Drug Delivery System) and reduction of the multi-resistant-bacteria emergence. It should be developed in the future, so the details on them are shown on Future work.

School-visits
Kanagawa prefectural Atsugi High School

The classes' goal was to raise students' interest and awareness of E. coli and to increase the knowledge about synthetic biology and gene ligation techniques.

The class got the result of an advance in knowledge and enhancement of the students' interest toward genetic modification (However, we could have got better results.)

So far, iGEM team Tokyo_Tech has "told the public about synthetic biology" in various ways with a familiar theme. However, we have not been able to influence all of the people. We thought that the reason relied on our way of communication. Therefore, we decided to ask a science communicator, who is an expert on telling science to the public.

National museum, nature and science, Tokyo
Science Communicator Mr. Ogawa

He told us that only representing the research cannot attract the audience's interest, and cannot get shrewd feedback. Also, he suggested that our story should have a clear conclusion that can tell people something.

We learned how to tell science to the public from a science communicator. From his talk, we noticed that we should design the whole project considering what the public will think after explaining our project.

In addition, we decided to make our project more attractive, through clarifying our project's goal and setting a clear conclusion.

Summary

We "went beyond the lab" and developed our project through human practice activities.

We have not just reflected opinions from dialogues with the public and experts about our project. Firstly, we investigated and discussed about the issues on our own. Secondly, in the light of the application of our project, we did experiments and modeling. Finally, we succeeded in the creation of "ACADwarfs" and "the growth and death wave by toxin-antitoxin on agar medium." Furthermore, we are doing experiments to make E. coli mutants secreting protein extracellularly. The products will probably lead to further developments of our project.

Achievements as mentioned above indicate that we have succeeded in the integrated Human Practice.

Suzukake Festival (May 14 & 15, 2016)

The Suzukake Festival, the school festival on the Suzukakedai campus, was held on May 14-15, 2016. We introduced iGEM and our projects to the visitors to let them know about synthetic biology and we received feedback.

Homecoming Day (May 21, 2016)

Homecoming Day is an annual event held at Tokyo Tech. We introduced iGEM and synthetic biology to the alumni. We realized that it is hard to explain our research themes and goals to the public clearly. Therefore, we decided to make videos about E. coli and genetic modification.

The remaking of the fairy tale "Snow White" by the Grimm Brothers has enjoyed a lot of versions since it was published in 1812 in the first edition of their collection Grimms" Fairy Tales. Some elements are common in any Snow White stories. However, because the authors add their original elements or features to the story, it becomes unique. We chose other two versions, Mirror Mirror" and "A Snow White Christmas" and designed their genetic circuits.