Team:GA State SW Jiaotong/Partnership
ShanghaiTech_China
MIBOM
This year, MIBOM decided to challenge the comminuted fractures and develop a new strategy for bone repair. MIBOM get inspiration from mussels, a creature that can adhere tightly to reefs through mussel protein in aqueous environment, and MIBOM designed a new biocompatible osteogenic glue.
HNOSS
In research published in the past year, two critical proteins, Gas6 and EGF, have shown promise for treating stress-related and androgenetic alopecia. Plasmids were constructed for transformation into Pichia pastoris, E.coli, and Bacillus subtilis respectively. Gas6 proteins will be purified from Pichia pastoris and EGF proteins will be expressed in E.coli using a T7 promoter to produce a cell-free protein treatment for hair loss.
06/19/21 Online meeting
GA_State_SW_Jiaotong and ShanghaiTech_China held an online meeting to discuss the cooperation on experimental and modeling design.
As for the experimental aspect, ShanghaiTech_China shared their previous experimental design of the toxin-antitoxin (TA) system, which includes a dual regulatory suicide system for inducing suicide in E.coli. Instead of using temperature changes as the condition that we intended, ShanghaiTech_China preferred to use tryptophan and lactose. As long as bacteria fall from the scalp to an environment without tryptophan and lactose, they will commit suicide.
Fig1.Two teams communicated about project, Hp and modeling.
As for the modeling design aspect, ShanghaiTech_China asked if our team could help design a model predicting high viscosity mucins. We suggested that, with a small amount of data, the prediction can be achieved by establishing equations based on biological properties of proteins like hydrophobicity, molecular weight, and ion charge. A partnership was established. As for the next stage, ShanghaiTech_China will provide us with some relevant information on the suicide system and the sequenced plasmid of suicide switch, with which our team will construct an appropriate Bacillus subtilis expression system. On the other hand, our team will help ShanghaiTech_China conduct their mucins model's preliminary design and construction.
07/19/21 In-person meeting
On July 17, GA_State_SW_Jiaotong and ShanghaiTech_China held an in-person meeting to exchange ideas on project cooperation. This meeting prepared for our future cooperation in this partnership.
ShanghaiTech_China proposed a solution regarding our question on how to construct the suicide switch. In the meeting, our team asked about the plasmid copy number, and ShanghaiTech_China answered that a homologous recombination method can be adopted for the construction of the suicide switch. At the same time, ShanghaiTech_China raised a concern that Bacillus subtilis has peripheral space that may cause the proteins unable to be secreted, which is a problem that we should look into.
Fig2.Two teams exchanged idea
As for the HP part, first, ShanghaiTech_China would help us find some experts whose expertise is in stem cells, protein drugs, and hair loss treatment. Secondly, both teams would establish their WeChat official accounts to introduce iGEM, different projects, and synthetic biology to the general public. The two teams would take turns to write articles about the most recent advancement in synthetic biology and jointly post the articles. In order to promote the educational influence of our projects, ShanghaTech_China will introduce our project to students in a rural area during their rural teaching project; and we will do some presentations in university, high school, and community in the United States to introduce both of our projects. Through mutual efforts, we hope that we could magnify our projects' influence and educational meaning in various areas.
Cooperation in Experiment
As for the experimental aspect, ShanghaiTech_China shared the design idea of the E.coli toxin-antitoxin (TA) system for GA_State_SW_Jiaotong. Unlike GA_State_SW_Jiaotong's previous design, which used temperature conditions to induce bacillus subtilis suicide, the TA system is a dual-modulated suicide system that uses tryptophan and IPTG to induce bacterial suicide. As soon as the bacteria leave the scalp and fall into an external environment free of tryptophan and IPTG, they will commit suicide.
In the In-person meeting in July, the two teams had in-depth discussions on the design of the suicide switch and the design of the protein viscosity prediction model. ShanghaiTech_China provided a more detailed design plan for GA_State_SW_Jiaotong and answered specific plasmid copy number and vector construction questions.
On September 27th, because the experimental conditions and operation of animal cell culture in GA_State_SW_Jiaotong are not mature, ShanghaiTech_China has excellent conditions and experience in this aspect. Therefore, GA_State_SW_Jiaotong transferred the efficacy experiment of EGF on HUVEC cells to ShanghaiTech_China.
Cooperation in Modeling
In terms of modeling design, ShanghaiTech_China solicited relevant design ideas from the modeling team of GA_State_SW_Jiaotong to predict mussel mucin's viscosity. GA_State_SW_Jiaotong suggested that, in the case of fewer related theorems, the viscosity prediction of proteins could be achieved by using a machine learning algorithm based on biological data such as hydrophobicity and molecular weight, and ion charge.
In the In-person meeting in July, ShanghaiTech_China asked the modeling team of GA_State_SW_Jiaotong another question demonstrating that hydrogels can be activated during compression by bone tissue healing. GA_State_SW_Jiaotong suggested that the microelement morphology of the hydrogel can be assumed, and then the real pressure of the hydrogel microelement can be calculated based on the stress deformation and the compression force relationship of the hydrogel. Finally, according to the obtained stress activation curve of the hydrogel, it can demonstrate that hydrogels can be activated during compression by bone tissue healing. ShanghaiTech_China thought this was a good idea, and they wanted to hand over the part to the modeling team of GA_State_SW_Jiaotong. In the end, GA_State_SW_Jiaotong finished it.
Hydrogel compression activation model
Cooperation in Human Practice
Student involvement fair
On September 14, we attended the Student Involvement Fair held by Georgia State University. It was a student organization fair where representatives from different organizations ranging from athletics to academic organizations came to advertise their clubs to GSU students and invite them to join their clubs. Together with Synthetic Biology Club members, we set up a stand with our posters to introduce the hair loss project and ShanghaiTech’s MIBOM program to the GSU students. Many students showed great interest in synthetic biology and signed up for the club. We also distributed flyers with a QR code for our survey on alopecia-related problems. Overall, the promotion was a great success.
Fig3.We introduced Hnoss and MIBOM.
Elementary school and junior high school
In late July, team members from ShanghaiTech_China went to Jiancao center to complete primary school, Yunlong County, Yunnan Province, China, to support teaching here. While supporting education, she also gave a lecture on synthetic biology to local elementary school and junior high school students. In addition to introducing synthetic biology and ShanghaiTech_China's project, she showed the students GA_State_SW_Jiaotong's project and PV. After the presentation, the students shared their opinions about our program. This event expanded the scope of our project. It can be the first time children in rural areas come across synthetic biology, which enriches their knowledge and may impact their future learning.
Fig4.MIBOM introduced Hnoss to Elementary school and junior high school
Lambert high school
On October 14th we organized a small presentation at Lambert High School. Lambert High School is a prominent High School situated in Suwanee, Georgia. We have been collaborating with them on their iGem project this year, and we got to visit and present our project to them as well as introduce our college lives in Atlanta, Georgia and in Chandu, China. The projects from our partnership team ShanghaiTech_China and NPU_China, and their college lives are also intoduced at the same time to make the students have more ideas about sythetic biology.
Lambert High School has been partaking in the iGEM competition for the last three years, so gaining their feedback along with insight into their project was very beneficial for us. We got to shed some light on how hair loss affects individuals, and how we were attempting to tackle it. At the end of our presentation we distributed a short survey to gather their opinion regarding hair loss as well. This provided us with an overall idea of how hair loss affects individuals in all age groups, so we would not be limiting our focus to the older populations alone.
Fig5.A small presentation with Lambert High School
10/16/21 Online meeting
On October 16, NPU_China jointly with GA_State_SW_Jiaotong and NWU_China gave an online presentation to about 30 high school students in China, who have experience in high school biology competitions and plan to continue studying biology in the future.
Fig6. An online presentation to about 30 high school students in China
First, students from NPU_China briefly introduced the development history and current status of iGEM and synthetic biology. Then Mr. Shang Xiaoya from the School of Life Sciences of Northwestern Polytechnical University conducted a sharing of biology and scientific research life with the theme of "From the New Coronavirus to Biochemistry". Finally, the NPU_China, GA_State_SW_Jiaotong and NWU_China three teams each introduced their own projects. While introducing our own project, we also introduced ShanghaiTech_China's MIBOM project and shared the university life for those high school students.
Through this event, we shared iGEM and synthetic biology related knowledge, and also promoted our project, which stimulated high school students' interest in natural sciences.
University
Southwest Jiaotong University
On September 24, we made a presentation about the iGEM projects of GA_State_SW_Jiaotong and ShanghaiTech_China to students and faculty from different majors in Southwest Jiaotong University. In this presentation, members of the GA_State_SW_Jiaotong team firstly gave an introduction to the project background and experimental design. Secondly, GA_State_SW_Jiaotong team members presented ShanghaiTech_China’s project in bone damage repair. This presentation expanded the popularity and influence of both projects.
In the process, students from different grades and majors of SWJTU actively gave feedback on the presentation. Later, they participated in a Q&A section, demonstrating a strong interest in our project and the iGEM competition, achieved the purpose of publicity.
Fig7.Posters and the presentation in SWJTU
ShanghaiTech University
On September 17, 2021, ShanghaiTech_China jointly hosted "Follow the iGEMers" with GA_State_SW_Jiaotong. The team leader Rui Su and other members jointly introduced synthetic biology and iGEM competition to about 100 students from various majors. From the content of synthetic biology to the style of iGEM, from experience to shareing, from biological science to computer science, ShanghaiTech_China team let more students from different disciplines realize that synthetic biology is a comprehensive discipline.
Fig8. "Follow the iGEMers"
In addition, we also introduced MIBOM project of ShanghaiTech_China team and Hnoss project of GA_State_SW_Jiaotong to the students. Most of the students are very interested in this, and a small number of students actively feedback to us: MIBOM can pay attention to some postoperative wound aesthetic problems caused by fracture surgery; Hnoss can take another look at the amount of alopecia gel applied.
Community
In China
On September 19th, GA_State_SW_Jiaotong conducted a community exhibition in Furong Community, Chengdu. Through exhibitions, basic knowledge about stress alopecia and bone injury treatment was introduced to the residents. After learning about our HNOSS project and ShanghaiTech_China’s MIBOM project, the residents scanned the QR codes to follow the WeChat official accounts of both teams and fill out the questionnaire. In the game session, we did some mini-games like memorizing cards and card flipping. Combining science with common knowledge, the games allowed residents to learn the knowledge of synthetic biology while having fun in the meantime.
Through community publicity, we successfully delivered the most advanced research results to the general public and get them to perceive the real-life application of synthetic biology.
Fig9.A community exhibition in Furong Community
In the United States
On October 10, GA_State_SW_Jiaotong team members came to downtown Atlanta to promote our project in the community. We put up our posters and distributed flyers on the streets, introduced synthetic biology and our project "Hnoss" to passing citizens, and many citizens showed interest in our project. At the same time, we also introduced the project "MIBOM" from ShanghaiTech_China, which promoted the promotion of the two parties' projects.
Through community publicity, we brought synthetic biology concepts and our programs to a larger population and connected them to the life of the general public.
Fig10. Promoted the Hnoss and MIBOM at downtown ATL
WeChat official account
To expand the influence of our project and the effect of popularization of science, we established a WeChat Official Account named "Hnoss 2021" in July 2021 and officially started operation in September, which has gained 389 followers, with a total of 6,030 views by October 17.
To achieve the purpose of project promotion and collaboration between teams, we introduced our project and forwarded other iGEM teams' projects on our official accounts, including ShanghaiTech_China and SJTang.
We also jointly created a topic named "Weekly Synbio" with ShanghaiTech_China to introduce the recent frontier progress of synthetic biology with simple and easy words once a week, and at the same time increase the knowledge of synthetic biology of our team members. We've published seven articles since we established this topic. The most clicked article has reached 3915 views; other articles also received a good response.
NPU_China
The Conference of China iGEMer Commu nity (CCiC), as the largest iGEM academic conference in China, attracts more than 100 teams from all over the country and more than 2,000 iGEMers to participate each year. In the annual iGEM competition, CCiC provides Chinese iGEMers a platform to communicate thoroughly.
We found that our project is very similar to the NPU_China team's project in the online meetup. After the first online meeting, we confirmed cooperation in partnership, experiment, HP, education, etc. The mutual help and interactive feedback between our two teams have played a significant role in promoting our projects during the process.
Project Design
Considering bio-safety and ethical issues, in the initial project design, both of us decided to use Bacillus subtilis as the chassis cell and apply a self-lysis system to control product release and bacterial reproduction to ensure safety
The NPU-CHINA team found in their research that the overexpression of Bacillus subtilis autolysin can be explored to lyse Bacillus subtilis.
In B. subtilis, some promoters only start transcription at the stable phase, such as PyqfD. In such a phase, promoters can be applied for the expression of specific products. By assembling the phase promoter of Bacillus subtilis and autolysin, a self-lysis system can be constructed.
The GA_State_SW_Jiaotong team proposed a bacterial toxin-antitoxin system that can regulate the growth of bacteria: the dual regulation system combining Lactose promotor and Tryptophan promotor with the sok-hok TA system. This controllable lysis system ensures that the live bacteria will not overproduce and pollute the environment during the application of the product.
Experiment
The NPU-CHINA team purchased the Bacillus subtilis WB600 strain at the beginning of the project and shared the strain with GA_STATE_SW_Jiaotong.
Fig11.BS WB600 single colony microscopy
Fig12.BS WB600 bacterial solution microscopy
Since our results in colony PCR and agarose gel electrophoresis experiments continued to be unsatisfactory. During online meetings, the NPU_CHINA team shared some tips on colony PCR and agarose gel electrophoresis, such as the appropriate water bath temperature for bacterial lysis and the appropriate amount of TAE buffer in running gel electrophoresis, and the ratio of sample to loading dye during sample spotting. In addition, the instructor of the NPU-CHINA team provided some competent cell preparation and transformation methods for Pichia pastoris and Bacillus subtilis, and NPU shared these protocols with the GA_STATE_SW_Jiaotong team.
The GA_STATE_SW_Jiaotong team successfully transformed Bacillus subtilis using the chemical transformation method provided by NPU-CHINA, which proved the protocol's effectiveness and provided the NPU-CHINA team with experience in operating details. Subsequent Bacillus subtilis transformation experiments were carried out smoothly for NPU_China.
Human Practice
Since the NPU-CHINA team did not participate in the 2019 and 2020 competitions, many team foundations need to be re-established and lack experience in iGEM competition. The GA_STATE_SW_JIAOTONG team provided expertise and guidance to the NPU-CHINA team in terms of Human Practice and education program design, which makes the NPU's project more diversified.
In the education section, the NPU-CHINA, the GA_STATE_SW_JIAOTONG, and the NWU-CHINA-A team carried out a synthetic biology presentation for high school students in China through online channels (Tencent meeting). This presentation explained some recent advancements in biology and briefly introduced synthetic biology. At the same time, the three teams shared their projects and related scientific knowledge. A total of 30 high school students participated in this presentation. They have experience in high school biology competitions and plan to continue studying biology in the future. After listening to this presentation, they showed significant interest in biological research.
For our public engagement in the United States, we visited Lambert High School in Suwanee, Georgia. We presented our project to them and briefly went over college life in Atlanta, Georgia, and Chandu, China. Lambert High School has been partaking in the iGEM competition for the last three years, so gaining their feedback and insight into their project was very beneficial for us. We shed some light on how hair loss affects individuals and how we were attempting to tackle it. At the end of our presentation, we distributed a short survey to gather their opinions regarding hair loss. The survey results present us with an overall situation of hair loss problems in all age groups, so we would not be limiting our focus to the older populations alone.
