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Making E.coli cells leading "self-destruction" after several rounds of division

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Pledged: 308,680 JPY
Target Amount: 280,000 JPY
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The inaugural year of iGEM Gunma

Inspired by synthetic biology, Mizuki Seta began from scratch to found the 2019 Gunma University iGEM team. She found motivated teammates from a variety of backgrounds interested in research and synthetic biology. In its inaugural year, iGEM Gunma aims to create a recombinant E. coli plasmid that can cause E. coli to automatically die after a controlled number of divisions. Part of the inspiration for this project came from a traditional Japanese phrase: “I curse you and all your descendants!” By creating a “cellular death” plasmid, iGEM Gunma aims to create a “curse” at the cellular level.

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miho otsuka

Discovering synthetic biology in a gym

Prior to entering Gunma University, I encountered synthetic biology for the first time while running in a gym. On a small display attached to a treadmill, I saw a TV program about synthetic biology. I had been looking for my life’s calling and sense of purpose, and after watching the program, I felt, “This is it!” Synthetic biology was very intellectually stimulating and interesting because it was completely different from the standard biology I had seen in high school. I watched the entire two-hour program (all while running on the treadmill) and soon after learned more about synthetic biology and iGEM.

iGEM is a huge international synthetic biology competition for students at the high school, undergraduate, and graduate levels. Over the course of a year, teams of ten students design and conduct synthetic biology experiments, concluding with a formal presentation of their research and experimentation in English at the iGEM Giant Jamboree. For those students going into scientific research, iGEM is an invaluable and formative opportunity. I decided to start an iGEM team at my university and was fortunate to find fellow teammates interested in the challenge of iGEM. Even though this is Gunma University’s first year participating in iGEM, our team has made great progress toward designing an E. coli death “curse.”

B1

Embedding controlled timed death into first-generation E. coli cells

Our theme builds on the project of the 2016 University of Tokyo team: making E. coli cells that change colors after every division. We came up with our theme in light of the fact that the iGEM competition includes two important criteria: improving previous projects and humor.

The 2016 University of Tokyo team successfully co-expressed distinct colors with each division by preparing different sets of genes for each generation to express. Both our current project and theirs focus on designing a function triggered by cell division. In our project, however, the function is significantly more complicated: E. coli are designed to die in a certain generation.

We established the theme of cellular “cursing” to make our project more original and humorous. Our theme comes from the traditional Japanese phrase, “I curse you and all your descendants!” We decided to program the slow-acting self-killing function into the first generation of E. coli, thereby “cursing” these cells. After several rounds of division, we could observe automatic death in daughter cells, representing the realization of the “curse.”

CcdB: molecular mechanism for cell death

We propose to exploit the cytotoxic CcdB protein as a means to lead E. coli to self-destruction. CcdB targets DNA gyrase, the topoisomerase that uses double-stranded breaks to undo the twisting of DNA, which is essential for replication. Inhibition of this critical function leads to failure in DNA replication and ultimately results in cell death.

How can we transfer the gene coding for CcdB into E. coli? It turns out that E. coli already have the CcdB gene. Usually, E. coli grow normally because they express CcdA, which inhibits CcdB. Thus, to promote self-killing of E. coli by CcdB, we plan to embed genes that code for CcdA inhibition, which would allow CcdB to become active.

B3

Background of our project: Idea for more comfortable guideline of GMO

We hope that our technology might one day simplify the regulation surrounding genetically modified organisms (GMOs). Many new technologies being developed in synthetic biology are highly controversial, ethically speaking, because of the implications they pose for editing of the genomes of many organisms. The Cartagena Protocol on Biosafety strictly regulates the handling of GMOs. To be clear, we are not against GMOs in general, but are concerned about the possibility that GMOs may negatively impact ecological diversity if they are released into the environment and outcompete native species. If we can set a “time bomb” in advance into a GMO, we can mitigate this potential environmental risk of GMOs.

B3

Support us

We are currently working on creating our self-killing E. coli cells and are trying to develop methods to count and distinguish among cells from different generations. Our proposed methods are so far based on those of the 2016 University of Tokyo team.

To successfully execute our vision from start to finish, iGEM Gunma requires around USD $18,000 to defray the costs of lab equipment and reagents, registration, travel, and lodging at the three-day Giant Jamboree in Boston. Despite the challenge of obtaining the necessary funds, we believe in the importance of our iGEM mission. Unfortunately, iGEM is not well known in Japan, so finding sponsors is not a straightforward task. But we know we can be successful with a crowdfunding approach, by garnering support from individual donors who believe in our mission. We plan to maintain a careful accounting of funds spent and keep our community of supporters informed of our future progress.

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Profile

I’m Mizuki Seta, leader of the iGEM team from Gunma University, Japan. Originally from Kanagawa prefecture, I had always dreamed to become a scientist since I was a young child. Currently, I am pursuing opportunities to gain intense, hands-on experience with research in synthetic biology. The iGEM competition challenges students to participate in the entire scientific process from start to finish: initial planning of a scientific project; gaining knowledge of background literature and knowledge; hands-on experimental work; and communicating results in writing and presentation. With a one-year competition season, iGEM is truly a marathon that requires dedication and perseverance. iGEM Gunma is committed to success.We hope we can count on your support and welcome all those interested in the mission of iGEM!

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Pledge Rewards

1,000 JPY (Tax not included)
Featured: iGEM2019 Report (pdf)
iGEM2019 Report (pdf)
1000 rev

We plan to share details of our progress, funny episodes throughout our research journey, and reflection of our first iGEM competition. We're counting on your support!

19 supporters are supporting with this reward. (Non quantity limit)

5,000 JPY (Tax not included)
Featured: RPG computer game of E.coli's adventure
RPG computer game of E.coli's adventure / iGEM2019 Report (pdf)
5000

A mutant E.coli goes on an adventure inside Gunma University Science&Engineering Campus!! We plan to share the game on the web for you to download. It will be 15-20 min long. We promise to make it fun and easy to play for children too! Join us and play the game!

7 supporters are supporting with this reward. (Non quantity limit)

10,000 JPY (Tax not included)
Featured: Invitation to the science cafe
Invitation to the science cafe / RPG computer game of E.coli's adventure / iGEM2019 Report (pdf)
10000

You will receive an invitation to our science cafe! If you wish, you may bring one friend to accompany you. It will be held in Gunma University Campus, Kiryu, Japan. Date to be announced soon. Please join us! In case you can't attend on the day, we will share the documents with you later. Transportation expenses are not included.

15 supporters are supporting with this reward. (Non quantity limit)

30,000 JPY (Tax not included)
Featured: Original team t-shirt
Original team t-shirt / Invitation to the science cafe / RPG computer game of E.coli's adventure / iGEM2019 Report (pdf)
R30000

We will send you our original team t-shirt, designed by us! It's the same one we will be wearing in Giant Jamboree of iGEM2019. Please note that this reward is shipped to those who have addresses in Japan.

0 supporters are supporting with this reward. (Non quantity limit)

50,000 JPY (Tax not included)
Featured: Your name in the Acknowledgement of our project
Your name in the Acknowledgement of our project / Original team t-shirt / Invitation to the science cafe / RPG computer game of E.coli's adventure / iGEM2019 Report (pdf)
30000

We plan to put your name in the Acknowledgement of our final project. Also we will share our project presentation slides with you. We count on your support!!

1 supporters are supporting with this reward. (Non quantity limit)

Supporters will be charged the funding amount only if the project reaches the funding goal (JPY 280,000) before 2019年05月20日 19時00分 (JST: GMT+9).
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1,000 JPY (tax excluded)

iGEM2019 Report (pdf)

1000 rev

19 supporters back
(Non quantity limit)

5,000 JPY (tax excluded)

RPG computer game of E.coli's adventure / iGEM2019 Report (pdf)

5000

7 supporters back
(Non quantity limit)

10,000 JPY (tax excluded)

Invitation to the science cafe / RPG computer game of E.coli's adventure / iGEM2019 Report (pdf)

10000

15 supporters back
(Non quantity limit)

30,000 JPY (tax excluded)

Original team t-shirt / Invitation to the science cafe / RPG computer game of E.coli's adventure / iGEM2019 Report (pdf)

R30000

0 supporters back
(Non quantity limit)

50,000 JPY (tax excluded)

Your name in the Acknowledgement of our project / Original team t-shirt / Invitation to the science cafe / RPG computer game of E.coli's adventure / iGEM2019 Report (pdf)

30000

1 supporters back
(Non quantity limit)

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