3.5
Team: PI gives the first introduction to iGEM competition.
On 5th March 2023 (Sun), PI Xin Hu held a talk that introduced the iGEM competition to students.
“Welcome to the exhilarating world of synthetic biology! This is a multi-disciplinary field of study
that involves the knowledge of biochemistry, computer science, mathematics, and even art design. The
iGEM competition provides numerous opportunities to engineering living organisms for various
purposes, for example green energy production and agricultural yield improvement. By joining our
school iGEM team, you may be able to work on cutting-edge projects and gain valuable hands-on
experience in the lab. Join us to shape the future with synthetic biology!” she said. Ultimately, 15
students successfully applied to join our big family!
3.6
Team: Team recruitment began.
Our PI Xin Hu set a series of different selection requirements to recruit different groups of
students who are passionate about synthetic biology.
3.23
Team: Our iGEM team CJUH-JLU-China is built.
Our team which includes groups of Wet lab, Model, Human practice, Wiki and Presentation called
CJUH-JLU-China was built.
4.9
Team meeting: Brainstorming identifies the disease detected by the project as breast
cancer.
4.12
Wet lab: miRNA is an important biomarker for cancer screening, and We decided to use
miRNA as a biological target for early screening of breast cancer.
4.15
HP: In our logo, "S" is inserted in the ⚡ middle, which stands for rapid screening
for breast cancer. The pink ribbon not only represents DNA molecules (in line with the theme of our
competition "synthetic biology"), but also shows our concern for breast cancer patients.
Wet lab: We
proposed the Nobel Prize-winning CRISPR/Cas9 technology, and used this as an entry point to discover
the CRISPR/Cas13a technology that is mostly used for detection, and carried out learning and
communication on the technology.
4.16
HP: Good action requires a clear timeline and action plan, so We discussed our proposed activities.
We initially worked out our mind map, which provided a direction for our actions.
4.17
HP: We sit together to discuss and write a timeline of IHP and Education.
Wet lab：We finally decided to use Leptotrichia wadei derived LwaCas13a protein by comparing Cas13a
proteins from several different sources, such as LwaCas13a and LbuCas13a.
4.20
HP: We create a team official in WeChat and an account on the Bilibili platform to get better
publication for synthetic biology and our project. To promote iGEM competition, we searched the
massive materials, finding that most of the existing videos revolved around topics like studying
abroad and scholarships, lacking in precision. Hence, we made a video to introduce iGEM competition.
Wet lab: We proposed that we can perform our test through fluorescence and lateral flow strips.
Model: We successfully performed differential expression analysis and preliminarily screened 9
miRNAs with high expression in breast cancer cells.
4.21
HP: To ensure that our programs are truly responsible for humanity, it is important hear the voice
of people from different communities. So We conducted a street interview to find out the public's
awareness of the early symptoms of breast cancer and the demand for breast cancer testing kits.
Model: We drew the ROC curve to narrow down the selecting range of miRNA from 9 miRNAs to 3 miRNAs.
4.23
Model: We innovatively use random forest models and SHAP analysis for feature importance analysis.
4.24
Model: We finished the survival analysis to verify miR-21-5p would play an important role in the
development of the breast cancer through Metabric and TCGA databases.
4.25
Model: We conducted literature research.
4.26
Team meeting.
4.28
Model: After reviewing the literature, we designed four crRNAs.
5.1
Model: We evaluated crRNA from three perspectives: the MFE, the GC content, the MFE of crRNA/miRNA
complex.
| crRNA |
MFE (kcal/mol) |
GC content |
MFE heterodimer (kcal/mol) |
| crRNA1 |
-9.8 |
0.39 |
-41.6 |
| crRNA2 |
-9.7 |
0.41 |
-40.9 |
| crRNA3 |
-9.6 |
0.41 |
-39.8 |
| crRNA full length |
-9.6 |
0.40 |
-42.5 |
5.6
HP: We released our first article on WeChat public account for the function of popularizing
synthetic biology. As soon as the article was published, it received a good number of views and
likes.
Model: Docking of crRNA and LwaCas13a protein was completed by HDOCK.
5.8
Team meeting
5.9
Model: Docking of LwaCas13a/crRNA/miRNA ternary complex was done.
5.11
HP: In order to understand the existing methods of breast cancer detection and the opinions of
medical professionals on our product, we interviewed Xing Hua, Director of Breast Surgery Department
of China-Japan Union Hospital of Jilin University. In this interview, it is revealed that breast
cancer patients heavily rely on routine examination. A low-cost, fast and convenient detection
solution is needed.
5.12
HP: After the initial experimental design of Brefast, we sought technical advice from Dr. Changlong
Song. Dr Song indeed appreciated the innovation and portability of Brefast, and he also suggested
ways to improve the sensitivity of Brefast when detecting the miRNA-21-5p level in patient’s blood
sample .
5.13
HP: We released our second article on WeChat public account to popularize the breast cancer .
5.14
Team meeting
5.15
HP: In speaking with Dr. Wang, we were pleased to hear that people's awareness of medical checkups
is increasing year by year, and that the percentage of early-stage breast cancer patients at her
hospital is growing. However, she believes that awareness of medical checkups and breast health
concerns is still not enough for the public. If our team can adopt more fun and accessible ways to
promote breast health, she believes it will be effective in raising awareness of breast health among
the general public, and in promoting Brefast.
5.16
Model: We finished molecular dynamics analysis of crRNA.
5.21
Team meeting
5.22
HP: We released our third article on WeChat public account to reporter the interview with Dr. Hua
Xing.
5.23
HP: We designed a brochure about breast cancer .
5.25
HP: We went to South Lake Park to perform an interview . In order to determine whether our early
breast cancer screening kit is really needed by the masses, CJUH-JLU-China team members Xinrui Wang,
Luohan Ge, Meng Yuan and Guanyu Huo carried out the first street interview in South Lake Park at
7:00 p.m. on May 24 around the masses' understanding of the early symptoms of breast cancer and
their attitude to the early screening kit.
5.27
Wet lab: We performed qRT-PCR experiments to verify the change in miR-21-5p expression in breast
epithelial cells MCF10A and breast cancer cells MCF7, demonstrating that miR-21-5p is highly
expressed in breast cancer cells.
Model: We finished Molecular dynamics analysis of LwaCas13a and crRNA/LwaCas13a complex.
5.28
Team meeting
Wet lab: We conducted the Transwell experiment to demonstrate that high expression of miR-21-5p can
improve the aggressiveness of breast cancer cells.
6.1
Wet lab: We collated the results of the wound healing experiment, which showed that the high
expression of miR-21-5p improved the migration ability of breast cancer cells.
6.2
HP: We Went to the Affiliated Middle School of Northeast Normal University for education work. To
demonstrate our project's commitment to high school students, under the guidance of Professor Wan,
our PI, we conducted a lecture on synthetic biology at Northeast Normal University Affiliated High
School.
6.3
HP: We released the forth WeChat public account.
6.4
Wet lab: We used SnapGene to build the pSB1C3-His-SUMO-LwaCas13a plasmid we needed to express the
LwaCas13a protein.
6.9
Wet lab: We cloned LacI OS, SUMO-LwaCas13a and pSB1C3 backbone sequences through PCR experiment.
6.13
HP: We Went to Changchun No. 1 Experimental Primary School for mission. Began by linking the
recently popular Spider-Man movies with the 2022 iGEM gold-winning project “Ocean Spider Silk”, we
used the analogy of building with blocks and assembling the puzzles.
6.17
Wet lab: We successfully conducted the golden gate assembly for pSB1C3-His-SUMO-LwaCas13a plasmid
and converted the assembled products into Trans1-T1.
6.18
Wet lab: We performed colony PCR to verify that plasmid assembly was successful.
6.20
Wet lab: We digested the assembled plasmid with single and double digestion to further verify its
assembly, and performed gel electrophoresis verification.
6.23
Wet lab: We demonstrated by SDS-PAGE gel electrophoresis that the addition of IPTG successfully
induced the expression of SUMO-LwaCas13a.
6.27
Wet lab: We performed SDS-PAGE gel electrophoresis on the supernatant and pellet obtained by
centrifugation after ultrasonic lysis of cells, and proved that SUMO-LwaCas13a mainly exists in the
lysed supernatant, and the addition of SUMO tag increased the solubility of LwaCas13a.
6.29
Wet lab: We demonstrated through western blot experiments that the expression of SUMO-LwaCas13a has
been successfully induced.
7.1
HP: We cooperated with the square dance club of Jilin University to promote health exercises.
CJUH-JLU-China team members Wang Xinrui, Ge Luohan, Yuan Meng and Huo Guanyu taught and demonstrated
the team's original breast exercises on the playground of Jilin University. The students were
enthusiastic and the breast health was spread in the jumping of the square dance.
7.2
Wet lab: We removed the SUMO tag at the front end of the LwaCas13a protein and completed the
LwaCas13a purification experiment. The entire expression purification process is shown on the gel.
7.4
HP: We designed board games. We also designed an English version of the game to make it easier for
people around the world to play.
7.6
Team meeting
7.8
HP: We participate CCiC conferences, exchange with other universities. We learn from each other and
our team is clearer about how to move the project forward.
7.10
HP: We communicated with DUT-China. We hold board games.To promote the awareness of breast health in
people, we held the Breast Cancer Awareness Game on 10th July 2023. Participants were divided into
five groups randomly. Each round had 4 participants who were given flag pieces.
7.16
Team meeting.
7.19
HP: We went to Long Quan community for education about breast cancer.To reach a wide range of people
with breast health knowledge, we organized a breast cancer prevention workshop at Longquan community
on July 19th. We began by introducing the prevalence of breast cancer and the risks factors. Then,
we taught the participants the public square dance for breast health.
7.23
Team meeting.
7.24
7.25
HP: We released the sixth WeChat public account.
7.26
HP: We communicated with AMFU-China.
7.27
Wet lab: We performed colony PCR to verify that plasmid assembly was successful.
8.2
HP: We published our first video on the Bilibili platform. Since square dance is one of the most
popular form of exercises in China, we collaborated with students majored in preventive medicine to
create a square dance with a set of movements to improve breast health. We chose the song ‘Dance
with Me’from movie Barbie as the background music of the dance, due to its relaxing melody and
energetic vibe.
Wet lab: We completed the purification experiment of crRNA.
8.4
HP: We publish our second video on the Bilibili platform. Our team created a promotional animation
featuring our project's mascot as the main character. There were both Chinese and English subtitles
to cater the needs of audience from different countries .
8.5
HP: We work with Tongji-software to create a brochure for the CRISPR/Cas system.
8.6
HP: We adapted the team promotional song and wrote commercial PPT and communicate with manager
Jingdong Wu.we interviewed Mr. Jingdong Wu, general manager of Jilin Guoke Innovation Plan
Investment Co, on August 6, 2023.
8.8
Model: We performed reaction kinetic function fitting.
8.13
Wet lab: We conducted fluorescence experiments look for the most reactive crRNA.
8.17
Wet lab: Fluorescence experiments explored the relationship between fluorescence intensity and miRNA
concentration.
8.20
HP: We released the seventh WeChat public account.
Wet lab: We conducted fluorescence experiments explore the optimal reaction concentration of
reporter.
Model: We performed reaction kinetic function fitting.
8.22
Wet lab: We wanted to make a test strip that could distinguish between high and low expression of
miRNA, so we designed its structure.
Model: The document of bioinformatics analysis finished . Our PI instructed us how to write and
revise in order to express our thoughts more clearly.
8.30
Wet lab: We performed lateral flow strips test successfully.
9.1
Wet lab: We designed the out shell of the lateral flow strips.
9.3
Team meeting
9.6
HP: We conducted a thorough market research and improved the quality of the product. We would like
to explore more possibilities of commercialization of BreFast with Mr. Wu, and went to his company
for a second interview.
9.9
Model: The document of Simulation and Verification of CRISPR/Cas13a System finished.
9.10
Wet lab: We finished the document of description. We worked together till deep night, and we all had
dinner in the office.
9.12
HP: We released the eighth WeChat public account.
9.13
HP: We went to Changchun No. 58 Middle School f12.r missionary education, aim to help the students
understand synthetic biology. We took the framework knowledge of biology as the entry point, and
talked about many biological phenomena such as the morphological structure of cells and ecological
environment homeostasis. We published our third video on the Bilibili platform.
9.16
Wet lab: We finished the document of Parts.
9.17
HP: We released the ninth WeChat public account.
9.19
Wet lab: We finished the document of engineering. He sun drew lots of pictures to make our
engineering page shine!
9.24
HP: We participated in the iGEM lecture of the fourth team of Jilin University.
9.26
Wet lab: The document of contribution finished.
9.29
HP: The document of IHP finished.
10.1
HP: The document of Education finished. Team members worked very late. Zhongyu took this photo
secretly.
10.6
HP: The document of collaboration finished. It was already 11o’clock at night, and the team members
were still discussing our project with the teacher on their way to leave office.
10.7
Wet lab: The document of results finished.
10.8
HP: We released the tenth WeChat public account.
