Design

Meeting the users’ demands is always the key of a successful product. Bearing this in mind, with our initial idea in developing miRNA inhibitors, we talked with the potential users of our product to gather their specific needs. Some interesting points that never came into our mind was raised in these interviews. For example, during the talk with Dr. Song Zhuo, the technical director of Genetalks, he talked about a few recently-published papers about cross-talks among different miRNAs. Building on these papers, he highlighted the potential needs of multi-targeting miRNA inhibitors and encouraged us to try crating one. For another example, during our interview with Professor Ping Li, she spoke highly of our dsDNA library based assembly system, and suggested us that a software assisting users to select correct nucleotides from the library might be necessary.

With these constructive suggestions, we adjusted our project design in the following aspects. Firstly, we tried assembling multiple-targeting miRNA inhibitors in our wet-lab experiments. Two miRNA Lockers, targeting two and four different miRNA molecules simultaneously were designed, assembled, and generated in our lab work and passed sequencing verification. Though cellular experiments on those Lockers have not been finished yet due to the time limits of iGEM, our current work have shown a great potential working in a multi-targeting manner.

Market demand:

Suggested by Dr. Song, we investigated existing microRNA inhibitors in the market. Our HQ team managed to contact with some companies to inquire prices of their miRNA inhibitors. According to the instruction given by those companies, we calculated the costs of each products. Then we found that prices of most miRNA inhibitors are quite high, to an extent of $52.68 for the transfection of one million cells. For most microRNA researching laboratories or microRNA antagonist therapies where large consumptions were needed, current approaches may be very costly. Thus, there is a big market for low-cost miRNA inhibitors.

The major reason for the high cost of current miRNA inhibitors was thought to be the highly costing chemical modifications like as in Locked Nucleic Acids (LNAs). For such matter, in our project design, unmodified DNA was directly used as miRNA inhibitors. Besides, due to the high cost of long-strand oligo DNA synthesis, we managed to develop a novel miRNA Locker generating procedure reducing the uses of long synthesized single strand oligo DNA, thus further reduce the cost.

Building on the modifications mentioned above, we recounted the cost of our scheme considering all reagent costs and experimental procedure. The cost transfecting one million cells using our scheme was successfully reduced to around $4.83. Together with the brilliant miRNA inhibiting effect of miRNA Lockers as well as its advantages on scalability, we thought our product could be a competitive candidate for miRNA loss of function researches as well as potential clinical uses.

Application prospect

Nowadays, miRNA inhibitors plays an important role in medical treatment. To know more information of miRNA antagonist therapy, we interviewed Professor Li LingProfessor and master’s supervisor of biochemistry and molecular biology department, school of Basic Medical Sciences, Southern Medical University, his main research direction is molecular mechanism of bone metastasis in tumor.

Professor Li Ling mentioned that the FDA program approved gene therapy technology and the first gene therapy was expected to be approved in January 2018. In addition, an RNAi (RNA interference) drug has entered the main clinical endpoint of phase three clinical trials. MicroRNA antagonist therapy is a promising new technique for treating diseases at gene level. With the wide application of this technology in clinic, many clinical drugs and therapies harmful to human body will disappear. However, new therapies will have their own advantages and shortcomings, old things cannot fade away immediately, and new things will also face many challenges to retain. Hospitals and clinicians should keep up with the breakthrough of medical technology, enrich the knowledge reserve of gene therapy technology and its treatment, and meet the new challenges at any time to serve the human health.

Safety

Realizing the importance of bio-safety issues, our HP team also talked with several professors about the possible biosafety concerns of our project. Though we were told that manipulating ssDNAs and use them as miRNA inhibitors ex vivo raises little safety concern, the other two aspects mentioned by the professionals raised our attention. On the one hand, Dr. Zhuo Song, the technical director of Genetalks noticed us that when used ex vivo, the potential toxicity of the miRNA Lockers may affect the normal function of the cells, on the other hand, Dr. Shuo Tu (Professor in the department of biochemistry and molecular biology, Nanchang University, mainly focus on tumor cell apoptosis) told us that if our miRNA Locker is capable of being used in vivo, its toxicity to the host animals as well as its pharmacokinetics should also be considered.

With their suggestions in mind, we made further adjustment on the experimental design of our project. Though with no specific data to show yet, we are preforming experiments determining the effect of our miRNA Lockers on the growth and apoptosis of the ex vivo cell cultures. Further in vivo experiments are still under planning and approving procedure of the ethical board of our university.

Idea Exchange

In the end of August, we attended FAFU_CCICFujian Agricultural and Forestry University Conference of China iGEAMer Community, and spent three days there.

The meetup is held for teams to share ideas and help each other. When communicating with other teams, we heard team BGIC-Union need plasmids which we designed a year before, then we decided to offer the plasmids, click here for details in collaboration page. In a few words, we had a fulfilling time in CCIC!

Exactly at this meeting, we found the project of team Lanzhou has some relevance to our project; their project is A novel method in controlling weeds and pests by tandem RNA Interference. Then we made communication in specific experiments at the venue.

After CCIC, we made deeper online interactionS with team Lanzhou with more team member participated.

we talked about each teams’ project mechanism, new situations encountered in experiments and progress of modeling and human practice.

Policy

Attentions on policy-making

MiRNA antagonist therapy is a brand new area that there is no policy to regulate it; however, we decided to grasp the trend of policy. As the product developer, we also shoulder the responsibility of advising the government when needed. Thus, we consulted Professor Li Ling and teaching assistant Tu Shuo on wht should be paid attention to when making a policy for miRNA antagonist therapy.

For new therapies and new technology, effectiveness and safety are two important indexes. In clinical application, there must be a strict regulation, including the indications and contraindications, and the patient's right to know. Hospital has the obligation to let patients fully understand the basic principle of anti microRNA therapy and the curative effect (cure rate) of new treatments’ individual differences, not every patient with antagonist treatment has good curative effect. The price should be transparent .

In the design of microRNA antagonist therapy, the hospital may need to carry out specifi gene sequence measurement for patients, and the relevant confidentiality rules should be signed in advance

Popularization

Publizing with new media

This year our track is foundational advance，and the product---miRNA inhibitors is not familiar to the public. Therefore, if we want to have the public engaged in our work, the very first thing is to let them make friends with miRNA and our project!

In order to achieve a greater influence and outreach, we decided to have our popular science article pushed by WeChat subscription (The WeChat ID of the subscription account is zaxue8, it aims to spread knowledge to the masses). In addition, we used many internet memes in this article for a more vivid expression, and it worked just as we expected!

By now, 2666 people have read this article, and the number is still increasing. Since it is on the internet, people can open it without limitation of time and space. And we can acquire the effectiveness of the publizing conveniently by reading rate and top comments.

You can read the article below as well as scaning the QR code by WeChat to read our articles on your phone:

Since we estimate miRNA is not popular among masses, we attached a small survey at the end of the article, and the result is:

This chart shows that the miRNA popularization still has a long way to go, and it might take a long time before taking the second step.

Do you want a picture book?

Many people, like me, will probably feel bored to read something contain nothing but words, especially when the words is about learning. I realized it after I entered college to find there are too few illustrations in textbooks. We do not want others to feel the same when they try to read our pamphlets, so we made it a beautiful picture book, printed and separated.

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Education

Lecture in high school

High school students are young, vibrant, and always open to new things.

Last year, our team had Lab open day for some high school students. One named Zhang Herui, showed great interest in iGEM competition and synthetic biology and was recruited into our team. This year, considering there are few references of miRNA mentioned in textbooks, we walked into her school to fill the blanks and introduce our project.

Moving our glow sticks for synthetic biology

In June, our team was invited to give a lecture on iTED Sharing held by the Squirrel Club of NUDT, the theme of the lecture is biotech 2.0-the synthetic biology and iGEM in my eyes. Through this lecture, we gave the present students an introduction of our research field. We aimed to share our ideas with undergraduates and get feedbacks form the interactive section. Also the popularization of synthetic biology is achieved.