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

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瀬田みずき
iGEM群馬大学チーム、代表
Pledged: 308,680 JPY
Target Amount: 280,000 JPY
Funded
110 %
Supporters
42
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Reached the funding target!

みなさま、こんにちは。iGEM群馬大学です。
いつも応援していただき、ありがとうございます!!!
iGEM2019のプロジェクトが完了しましたので、ご報告をさせていただきます。
結果から申しますと、弊チームは無事ボストンで開催されたGiant Jamboreeに参加し、銅メダルを獲得いたしました!!
リーダーの北を含む3名のメンバーが代表として、10/30~11/5の約1週間ボストンに滞在しました。
ボストンは群馬よりも寒く、気温が2℃~12℃くらいの日もありました。
Giant Jamboreeは世界中のiGEMチームが成果を発表する場所です。
Hynes Convention Centerという大きな建物内で開催され、5000人以上の学生たちが集結しました。
私たちは10/31にチームとしてのチェック・インをすませ、翌日の11/1にプレゼン+質疑応答と2回のポスターセッションを行いました。使用言語はすべて英語です。
緊張しましたが、自分たちの発表をたくさんの人が注目して聞いてくださるという状況には胸が高鳴りました。
今年度、私たちはBL21(DE3)pLysSという株の大腸菌を溶菌しやすくする遺伝子パーツをデザインしました。
iGEMでメダルを獲得するには、それぞれ設けられた条件を満たす必要があります。
私たちに起こった問題は、pLysSと私たちオリジナルのパーツを載せたプラスミドバックボーンの抗生物質に対する耐性はどちらもクロラムフェニコールに対するものであることでした。大腸菌に望みの遺伝子が導入されたかどうかは、抗生物質(耐性を持たない大腸菌を殺す物質)入りの培地で全ての大腸菌を培養して確認します。つまりそのままでは両方のプラスミドをもつ大腸菌を選別することができないので、私たちはオリジナルパーツを別のプラスミドに移そうとしました。
しかしこの実験に予測より何倍も時間がかかったため、残念ながら時間切れとなり金、銀の条件を満たすことは結果的にかないませんでした。
今年は例年に比べて金メダルを取ったチームの数も多く、世界中から集まった他のチームのプロジェクトの完成度に驚かされることも多くありました。ですが、私たちは初年度で銅メダルという結果をきちんと残すことができたことを誇りに思います。
数多の障壁を乗り越え、最後までたどり着くことができたのも、温かく支えてくださったみなさまのおかげです。心から感謝申し上げます。
より詳しいご報告については、現在大会レポートを作成中ですので今しばらくお待ちください。
こちらは弊チームのチームwikiです(英語)。
もしよろしければこちらもぜひご覧ください。
https://2019.igem.org/Team:Gunma
改めまして、弊チームをお見守りいただき、また応援していただき本当にありがとうございました。
来年以降もできることならばチームを存続させ、いずれ銀、金メダルを獲得したいと考えておりますが、関係者各位とよく相談した上で今後の進路を決めていければと思っております。
ただいま各種お礼の準備が進行中ですので、続報をお待ちいただければ幸いです。
iGEMの公式ページはこちら:https://2019.igem.org/Main_Page

Comment from academist staff
The inaugural year of iGEM Gunma

miho otsuka

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.

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.”

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.

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.

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.

Profile

瀬田みずき

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!

Project timeline

Date Plans
2019年1月 iGEM申し込み
2019年3月 クラウドファンディング挑戦
2019年2月〜10月 実験(試薬購入)
2019年4月 参加費支払い
2019年10月 個人登録費(8万円×渡航者人数)支払い
2019年10月31日〜11月4日 ボストンにて発表

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Featured : iGEM2019 Report (pdf)

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!

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iGEM2019 Report (pdf)

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Featured : RPG computer game of E.coli's adventure

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!

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Featured : Invitation to the science cafe

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.

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Featured : Original team t-shirt

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.

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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!!

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Supporters will be charged the funding amount only if the project reaches the funding goal (JPY 280,000) before 19:00 on May 20, 2019 (JST: GMT+9).
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RPG computer game of E.coli's adventure and others

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Invitation to the science cafe and others

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33,000 JPY(tax included)

Original team t-shirt and others

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55,000 JPY(tax included)

Your name in the Acknowledgement of our project and others

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