Team:Shanghai Metro/Implementation

Shanghai_Metro

Implementation
Background
Our team is working on a patented strain protection technology based on type VI secretion system effectors. A conversation inspired us with a biology professor. We learned about the wide range of uses of strains and the lack of patent protection for strains in the biomedical industry today. After deciding on the topic, we started our market research, which consisted of a questionnaire, field research, and interviews with experts. We first created a questionnaire with contents related to patent protection and publicized it on WeChat, Weibo, etc., as a way to reflect people's awareness of patent protection. Since there were some limitations in answering the questionnaire and the people who responded were relatives and friends, we conducted another field research. We asked people of different professions in nearby shopping malls to see what the public knew about the strain and their thoughts about the technology. Finally, we invited some experts and interviewed them, among them biologists and lawyers. We realized the inadequacy of patent protection laws in biomedicine and the frequency of strain theft from our conversations. As a result, we became more sure of the benefits of this topic and gained a deeper understanding of it.
Product objectives and user groups
Our product is mainly designed to protect the strain patents of biomedical experts. The goal is to make encryption for the invented microorganisms so that the stolen strains cannot be used. Secondly, it will allow the encryption system to leave a certain mark on the strains and facilitate the subsequent legal intellectual property disputes involved, as this technology can be used as legally recognized evidence. The strain patent protection technology user group is mainly people or companies that have invented or researched strain patents.
Product introduction
Figure 1. Workflow of the product
Our design was inspired by the Type VI secretion system (T6SS) of Pseudomonas putida. T6SS kill eukaryotic predators or its competitors by injecting anti-bacterial effectors, the toxic substances, into eukaryotic or prokaryotic cells. Pseudomonas putida itself will not be killed by the anti-bacterial effectors because it secrets immunity proteins specific to each of their antibacterial effector proteins to protect itself.
In order to protect the bacterial strains of the customer company, we need to transfer the anti-bacterial effectors tke2/4 and the immunity effectors ike2/4 into the plasmid of the bacterial strains. The expression of immune effectors needs to be induced by tetracycline, a type of inducible promoter, in the appropriate concentration. Thus, we sell the recipe to the company to protect their bacterial species.
Recipe:
1) Pus232-ike2/4: double restriction enzyme sites of Pus232 and their complementary DNA sequence added to the two ends to ike2/4.
2) Pus232-ike2/4-tke2/4: the design of single restriction enzyme site of Pus232-ike2/4 and its corresponding homologous recombination enzyme.
3) Optimal tetracycline concentration: 0.01ug/mL.
Product functionality
We have conducted several functional tests to analyze the performance of our design. As seen above, the control group Pus232 DH5α yield blu strains but the experimental group Pus232-ike4-tke4 had no blue strains, which could indicate that our design worked in DH5α.
In addition, we also determined the suggested concentration range of tetracycline for the experimental bacterial species growth by measuring the growth curves under tetracycline with different concentrations cultivate with comparing to the control group and it turn out to be 300~500ug/L.
Marketing plan and product selling points
Our main approach to marketing and promotion is to publish papers related to this technology in major biology journals so that biomedical scholars can see such a technology. We will also go to different large biomedical companies to promote it and present our research results at various large companies. In addition to this, we will also find famous researchers and corporate executives in the biomedical community to help us promote our research, using their popularity and influence to help us promote our research. Our marketing will be done both online and offline to promote the technology in the broadest way. At the same time, we position the product in the niche market to better help researchers who need this technology and give them a more tailored effect while spending the least money to achieve the best publicity.
The main selling point of our product is to protect researchers, protect their inventions and creations, and help them save time and cost on subsequent legal disputes. Because so far, China's Intellectual Property Law is still not perfect, especially in biomedicine, so in case of strain patent theft, it will cost a lot of money and time in legal matters. Therefore, our technology saves them a lot of unnecessary money and trouble. In addition, our product has another selling point, that is, we are the first team in China to research this technology, and we are at the forefront of this industry. We are also the pioneer of strain patent protection technology, so our technology can break the current problem of strain theft, and also considered to promote the development of the patent protection industry, and also to reflect the importance of patent protection, and mobilize people. It also demonstrates the importance of patent protection and raises awareness of it.
Future challenges and potential risks
1) The recipe of tetracycline is at risk of being stolen. If the recipe is stolen, will there be other induced promoters to replace tetracycline? Regarding this hidden danger, we conclude that the bacterial strains that different customers need to protect are different. Therefore, the sensitivity of tetracycline is different for different bacterial strains. That is to say, the results of tetracycline may be uneven for different customers.
2) Recently, we have only transfer the Pus232-ike2/4-tke2/4 to the plasmid of E.coli. Thus, we can only be sure that our technology can be used to protect E.coli. We need to explore more about whether our product can be used to protect other targeted bacterial species in the future.
Bibliography
1. Hernandez, Ruth E., et al. “Type Vi Secretion System Effector Proteins: Effective Weapons for Bacterial Competitiveness.” Cellular Microbiology, vol. 22, no. 9, 2020, doi:10.1111/cmi.13241.
2. Rossi, R. “Functional Characterization of the T4 DNA Ligase: A New Insight into the Mechanism of Action.” Nucleic Acids Research, vol. 25, no. 11, 1997, pp. 2106–2113., doi:10.1093/nar/25.11.2106.
3. “Homologous Recombination in Procaryotes.” Microbiological Reviews, vol. 52, no. 2, 1988, pp. 304–304., doi:10.1128/mr.52.2.304-304.1988.