iGEM 2017 — Team Shenzhen_SFLS

[![](https://static.igem.org/mediawiki/2017/8/8b/HQ_page_logo.jpg)](https://2017.igem.org)

# Team:Shenzhen SFLS/Notebook

Team:Shenzhen SFLS/Demonstrate - 2017.igem.org

[![Shenzhen_SFLS](https://static.igem.org/mediawiki/2017/7/73/Sfls-logo.gif)](https://2017.igem.org/Team%3AShenzhen_SFLS)

* [Project](https://2017.igem.org/Team%3AShenzhen_SFLS)
  + [Description](https://2017.igem.org/Team%3AShenzhen_SFLS/Description%20)
  + [Design](https://2017.igem.org/Team%3AShenzhen_SFLS/Design%20)
  + [Experiment](https://2017.igem.org/Team%3AShenzhen_SFLS/Experiments%20)
  + [Demonstrate](https://2017.igem.org/Team%3AShenzhen_SFLS/Demonstrate%20)
  + [Collaboration](https://2017.igem.org/Team%3AShenzhen_SFLS/Collaborations%20)
  + [Achievement](https://2017.igem.org/Team%3AShenzhen_SFLS/Achievement%20)
* Parts
  + [basic part](https://2017.igem.org/Team%3AShenzhen_SFLS/Basic_Part%20)
  + [composite part](https://2017.igem.org/Team%3AShenzhen_SFLS/Composite_Part%20)
* [Modeling](https://2017.igem.org/Team%3AShenzhen_SFLS/Model)
* Human Practice
  + [GOLD](https://2017.igem.org/Team%3AShenzhen_SFLS/HP/Gold_Integrated%20)
  + [SILVER](https://2017.igem.org/Team%3AShenzhen_SFLS/HP/Silver%20)
  + [Publicity](https://2017.igem.org/Team%3AShenzhen_SFLS/Publicity%20)
  + [Biosafety](https://2017.igem.org/Team%3AShenzhen_SFLS/Biosafety%20)
* [Notebook](https://2017.igem.org/Team%3AShenzhen_SFLS/Notebook%20)
  + [Notebook](https://2017.igem.org/Team%3AShenzhen_SFLS/Notebook%20)
  + [protocols](https://2017.igem.org/Team%3AShenzhen_SFLS/Protocols%20)
* [Team](https://2017.igem.org/Team%3AShenzhen_SFLS/Team)
  + [About us](https://2017.igem.org/Team%3AShenzhen_SFLS/Team)
  + [Attributions](https://2017.igem.org/Team%3AShenzhen_SFLS/Attributions%20)

Team:Shenzhen SFLS/Protocol - 2017.igem.org

# Notebook

[December, 2016](#index_1 "1")
[January, 2017](#index_2 "2")
[February, 2017](#index_3 "3")
[March, 2017](#index_4 "4")
[April, 2017](#index_5 "5")
[May, 2017](#index_6 "6")
[June, 2017](#index_7 "7")
[July, 2017](#index_8 "8")
[August, 2017](#index_9 "9")
[September, 2017](#index_10 "10")
[October, 2017](#index_11 "11")
[November, 2017](#index_12 "12")
回到顶部

#### December, 2016     DETAILS

At the end of 2016, our team was founded.

We sent mails to foreign teams and got in touch with them.

#### January, 2017     DETAILS

##### Theory Experiment

After our first group meeting, we were divided into 4 groups and got the first assignment since we join the team — read as much wikis as we can and came up with embryonic form of the new project during the winter holiday.

From 21st to 26th, our team members held a meeting to report the wikis we have read before and to communicate our ideas about the project. We also learned some skills that we should have in the laboratory.

##### Human Practice

Raised plans for future work to get sponsorship.

Ran our official account on Wechat and published the first article on 31st January .

#### February, 2017     DETAILS

##### Theory Experiment

In this month, we submitted our registrations and got the qualifications of attending the competition.We spent the whole February reading wiki and completing the basement of our projects.

Published the second article to popularize basic knowledge of biology on Wechat official account on 19th February .

In the late February, we had a discussion about choosing a project. Here are our early projects:

- Biosynthesis Resveratrol
- Treating Trichophyton rubrum infection by Biosynthesis hBD3 (human Beta-Defensin 3)
- Treating hyperplastic scar by Biosynthesis MMP-3 (matrix metalloproteinase 3).
- Detect dangerous gases’ concentration in mines by Remold E.coil.
- Utilizing remold E.coil to take in harmful gases
- Cancer treatment.

Finally, we chose the project 2 —Treating Trichophyton rubrum infection by Biosynthesis hBD3 (human Beta-Defensin 3)

##### Human Practice

Published the second article to popularize basic knowledge of biology on Wechat official account on February 19th.

#### March, 2017     DETAILS

##### Theory Experiment

During the group meeting in March 8th, we came up with several more directions to treat Trichophyton rubrum infection.

Here are our tentative ideas:

- Improve those inhibitors that are normal but imperfect now are being used;
- Send hBD3 into human body and complete the treatment;
- Inhibit keratinase that Trichophyton rubrum create, inhibit the immune evasion of Trichophyton rubrum or reduce the drug resistance.

##### Human Practice

Published the third article about transgenosis and transgenic products on Wechat official account on March 24th.

#### April, 2017     DETAILS

##### Theory Experiment

In this month, we were going to express hBD3 in the periplasmic space of E. coli at first, but after a period of study, we think that’s an unfeasible way.

On April 22, we went to the Laboratory of Molecular Biology, Shenzhen PKU-HKUST Medical Center and took some experimental training. On April 25, we met Dr. Haoqian Zhang, a professor in Chinese Academy of Sciences, and discussed about our
project. He gave us much advice.

Being inspired by the instructors, we found some serious problems that may affect our future study, for example: 1) How to avoid the formation of inclusion bodies? 2) How can we make sure the molecular chaperones service can work as expected?
3) The feasibility of the project is actually low. What can we do?

There were also many problems found in Human Practice, like: 1) How to dispel the discrimination of that disease in public? 2) How to make the medicine more convenient for people to use? 3) How to shorter the time for treatment?

##### Human Practice

On April 1st, we released a questionnaire about Trichophyton rubrum.

On April 20th, we made a wonderful speech about our project and synthetic biology at Scientific Festival in Shenzhen Foreign Languages School.

Opened an account on Twitter and then published our articles on both Wechat and Twitter.

On April 29th, we met up with universities team and high school teams from south eastern China at Shenzhen University.

On April 30th, we closed the website of Trichophyton rubrum questionnaire.

#### May, 2017     DETAILS

##### Theory Experiment

We prepared for the experiment of Trichophyton rubrum in May.

We ordered the germination conidia of Trichophyton rubrum and did a bacteriostatic effect test.

We found that the secretion of hBD-3 using hBD-3 ELISA was not detectable.

We did some pre-experiment and wrote an experiment report.

However, through human practice and meet-ups, we found that the project based on Trichophyton rubrum was too difficult for us to do, so we started to look for other possible directions.

We found another aspect that we could do thing to improve it: cancer.

##### Human Practice

Prepared souvenirs and related products about our project.

Planned to organize a meet up with Shenzhen College Of International Education in summer vacation and sent out the invitations.

#### June, 2017     DETAILS

##### Theory Experiment

Through reading articles we decided to use CRISPR/Cas9 System to treat cancer.

From June 7, we started to study about CRISPR/Cas9 system.

In this month, we submitted the safety form.

##### Human Practice

Released a questionnaire about cancer on June 3rd.

On June 5th, w delivered a speech about our project in Shenzhen Foreign Language School(Junior High).

Kept on searching background information of the two projects and decided which project to choose.

On June 11th, we interviewed Dr.Jinfen Zhen from dermatological department in the hospital of Chronic Disease Prevention and Treatment in Luohu district in Shenzhen.

On June 19th, we interviewed Dr.Boran Cheng from oncology department in PUSH.

On June 30th, we closed the website of cancer questionnaire.

#### July, 2017     DETAILS

##### Theory Experiment

After the human practice part, we had a profound discussion about changing the project. Finally, we changed our project into the treatment for cancer in this month.

On July 5, we designed our Single-Guide RNA (sgRNA) which could specifically bind to the mutant BRAF gene.

On July 8, we started to culture the melanoma cell lines A375 and G361.

On July 9, we cultured the bacteria containing our plasmid containing our sgRNA and Cas9 protein.

On July 11, we extracted genome from A375 and G361 cell lines to check if the cell lines had BRAF V600E mutation. We extracted the plasmids to prepare for cell transfection.

On July 13, we transfected the cells with our plasmid.

On July 14, we assigned the task of writing different parts of our wiki to everyone and started writing our abstract, description and design of our wiki.

We cultured the cell lines.

On July 20-21, we submitted 4 parts.

On July 26, we cultured the cells in a 6-well plate and transfected the cells in order to measure cell migration by wound healing assay.

On July 27, we seeded the cells in a 96-well plate and trasnfected the cells in order to measure the cell proliferation by Transdetect Cell Counting Kit (CCK). We planned to scratched wounds across the cells but failed, for G361 cells didn’t
cover the dish.

On July 28-30, we stained the cells to detect the results of CCK assay. We cultured the cells.

On July 31, we finished writing our first draft of our abstract, description and design.

##### Human Practice

Attended the 19th International Botanical Congress.

On July 10th, we met up with Southern University of Science and Technology iGEM team.

From July 24th to July 30th, we went to Bali island to teach teenager “How to keep healthy”.

On July 30th, we delivered a public speech about iGEM, Synthetic Biology and Melanoma in Shenzhen Bookcity and gave out questionnaires while the speech was going on.

#### August, 2017     DETAILS

##### Theory Experiment

On Aug 1, we exchanged the Cell medium. We froze the Cells. We seeded the cells in a 6-well plate and transfected the cells to measure the cell migration by Transwell assay. We seeded the cells in a 96-well plate and transfected the cells
in order to measure the cell proliferation.

On Aug 2, we stained the cells to detect the results of CCK assay. We stained the cells to detect the results of Transwell assay.

On Aug 3, we subcultured the melanoma cell, exchanged the cell medium and stained the cells to detect the results of CCK assay.

On Aug 4 – 5, we stained the cells to detect the results of CCK assay and subcultured the melanoma cells.

On Aug 8, we seeded the cells in a 6-well plate and transfected the cells to measure the cell migration. We cultured the bacteria containing our plasmid for plasmid amplification.

On Aug 9, we extracted the genomic DNA of the cells. We transfected the cells.

On Aug 10-12, we seeded the cells in a 6-well plate and transfected the cells for TA cloning assay.

On Aug13, we seeded the cells in two 6-well plate and transfected the cells for cell apoptosis and migration assay.

On Aug 14, we transfect the cells, cultured the bacteria containing our plasmid for amplification, stained the cells to detect the results of CCK assay and flow cytometry assay.

On Aug.15, we extracted the plasmids and stained the cells to detect the results of Transwell assay.

On Aug19, we attended the meet up and introduced our project.

In the whole August, we gained a lot from experiments.

##### Human Practice

From Aug 19th to Aug 20th, we cooperated with Shenzhen College of International Education to organize a meet up with several iGEM teams.
Completed the Attribution part.

Published articles about biology on Wechat.

#### September, 2017     DETAILS

##### Theory Experiment

We started to write the experiment and demonstrate of our wiki and the preparation of confirmatory experiment and presentation in the Giant Jamboree.

On Sept 29, we submitted some parts.

##### Human Practice

Searched information about Biosafety and preliminarily met the requirements of iGEM headquarter.

Started to edit the Human Practice part of our wiki.

Attended the Clubs Day of SFLS and Introduced iGEM competition and our project to our schoolmates.

Searched for information about CRISPR related safety and ethic issues Gave out questionnaires to survey if the public pay plenty attention to melanoma and their attitudes towards it.

#### October, 2017     DETAILS

##### Theory Experiment

In the whole month, we were polishing our wikis and preparing for the Giant Jamboree.

We also communicated with the iGEM team BGIC\_Union.

##### Human Practice

Interviewed experts about future work and their views on our project

Finished & Arranged wikis

Finished the Human Practice part of poster

Delivered a speech in Baihe Foreign Language

Arranged & Analyzed the result of our CRISPR questionnaire and published articles about the result

Raised a fund raising on the Internet

Prepared speech and PPT for presentation

Completed the single prize application in judging form

Made the biosafety video

#### November, 2017     DETAILS

Attend the Giant Jamboree!!!!

#

### [SFLS.](home.html)

Shenzhen Foreign Language School, Yantian Road No.1, Yantian District, Shenzhen, Guangdong, China

* [Home](https://2017.igem.org/Team%3AShenzhen_SFLS)
* [Description](https://2017.igem.org/Team%3AShenzhen_SFLS/Description%20)
* [Design](https://2017.igem.org/Team%3AShenzhen_SFLS/Design%20)
* [Experiment](https://2017.igem.org/Team%3AShenzhen_SFLS/Experiments%20)
* [Demonstrate](https://2017.igem.org/Team%3AShenzhen_SFLS/Demonstrate%20)
* [Collaboration](https://2017.igem.org/Team%3AShenzhen_SFLS/Collaborations)
* [Achievement](https://2017.igem.org/Team%3AShenzhen_SFLS/Achievement)

* [Basic Part](https://2017.igem.org/Team%3AShenzhen_SFLS/Basic_Part%20)
* [Composite Part](https://2017.igem.org/Team%3AShenzhen_SFLS/Composite_Part%20)
* [Modeling](https://2017.igem.org/Team%3AShenzhen_SFLS/Model%20)
* [HP-Silver](https://2017.igem.org/Team%3AShenzhen_SFLS/HP/Silver%20)
* [HP-Gold](https://2017.igem.org/Team%3AShenzhen_SFLS/HP/Gold_Integrated%20)
* [Biosafety](https://2017.igem.org/Team%3AShenzhen_SFLS/Biosafety%20)
* [Protocols](https://2017.igem.org/Team%3AShenzhen_SFLS/Protocols)

* [About us](https://2017.igem.org/Team%3AShenzhen_SFLS/Team%20)
* [Attribution](https://2017.igem.org/Team%3AShenzhen_SFLS/Attributions%20)
* [Engagement](https://2017.igem.org/Team%3AShenzhen_SFLS/Engagement)
* [Contribution](https://2017.igem.org/Team%3AShenzhen_SFLS/Contribution)
* [Notebook](https://2017.igem.org/Team%3AShenzhen_SFLS/Notebook%20)
* [Publicity](https://2017.igem.org/Team%3AShenzhen_SFLS/Publicity%20)
* [Improve](https://2017.igem.org/Team%3AShenzhen_SFLS/Improve)

* [![](https://static.igem.org/mediawiki/2017/6/6c/Sfls_twitter_before.svg)](https://twitter.com/SFLS_iGEM)

[![](https://static.igem.org/mediawiki/2017/b/bf/HQ_menu_logo.jpg)](https://2017.igem.org)