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项目简介……（蚊子传播的疾病，引起多少人的死亡etc；1-2句话）

但是我们的产品将要如何进入市场，如何很好的贴合人们对杀蚊产品的需求，如何给社会带来帮助与贡献？在项目进行的过程中，我们逐渐意识到了仅仅靠AIS-China队伍里队员们的想法来建立产品是远远不够的，很容易出现市场定位错误的情况。

因此我们通过问卷调查和华南交流会、CCIC大会等交流活动了解到群众对novel杀蚊产品的需求，走访了CDC officials、公园工作人员、宠物医生等专家，和在shRNA农药上颇有了解的2024 SZU-China，以及致力于RNA检测的2024 Tsinghua-M进行了线上交流，以确定产品后续的发展方向，并确认了产品的市场竞争力。在了解到citizens对安全性的concern后，我们seek perpectives of ……（safety）on RNAi mosquitosicide and 活菌。为了引起他们对mosquito control的重视，我们还从社区和学校入手，给public和pupils普及蚊子会带来的疾病。

我们的HP的目标是希望能整合所有受到目前灭蚊产品弊端困扰以及致力于mosquito control人群，开展与experts from various fields的交流，以促进Moskilla to be as good and responsible as possible to enter the real world。

We are committed to involving human practices in the engineering design process to successfully meet the needs of our potential end-users. We believe that for new technology to be accepted by society and enter the market, it is essential to listen to, evaluate, and incorporate the voices and backgrounds of potential end-users into the design. This is known as consumer-driven product design. In addition to the voices of potential end-users, we also want to value the voices of society, as this will help promote Moskilla to be as good and responsible as possible when entering the real world. So we define the users and related social groups involved in the project as our stakeholders.

For every stakeholder that has an influence on the development of Moskilla, we have prioritized our core values, including public health, profitability, environmental safety, innovation and privacy. We strive to integrate these values into Moskilla, helping us to identify consistency and resolve conflicts among them.

You can click on each value to learn more about our efforts to implement them.

Public health

Environmental safety

Innovation

Profitability

Privacy

Value title

Value quote Value Cite

Value paragraph

∙ Reflection on AREA framework

To confirm the impact of a project, it is necessary to engage in effective two-way communication with various stakeholders. Therefore, we need a structured tool to help us integrate feedback received from stakeholders and make timely adjustments to project design based on feedback. Inspired by 2022 TU-Eindhoven, we use the four steps of their optimized Area framework as guidance to help us engage in targeted communication.

Reflecting on our project, we define the purpose of the communication event. Keeping the purpose in mind, we gain feedback while engaging with our stakeholders, take actions on the implementation of our project, and outline our next steps for further anticipation. By following this cycle, we have advanced our communications with stakeholders throughout the journey, ultimately achieving one milestone after another.

Purpose after reflection: This section shows the goals we set before we start our IHP activities, which is at the heart of every event. This ensures that we won't be distracted by irrelevant voices or ignore the current needs of specific stakeholders. Also, this demonstrates what drives our human practices journey to a milestone.

Gains by engagement: This presents the concerns and suggestions expressed by stakeholders. Under our values, this is all about ensuring that the other party agrees with the accurate information.

Implementation by actions: This shows our actions. Based on the information given by stakeholders, what kind of changes our projects proactively make to address stakeholder concerns.

Outlook for anticipation: This shows our next step in action and is what drives our human practices journey to a milestone. The interests of stakeholders from different perspectives may be conflicting, and what we should do next is to integrate our values into Moskilla, helping us to identify consistency and resolve conflicts among them.

∙ Events category

We have consulted with experts from various fields, who have provided valuable insights from multiple perspectives. Their contributions have been instrumental in identifying issues and formulating our solutions, and have also helped us understand the diverse impacts that Moskilla might cause.

After analyzing their characteristics and their interests in our project, we divided their outreach events into the following 5 categories based on their contributions to the development of Moskilla, the need, implementation, safety, laws and regulations, business.

You can click on each event to learn more about our definition of corresponding events.

The need

This section analyzes the reason why specific stakeholders need our project and elaborates how our project helps deal with their problems combining with the suggestions from experts.

Safety

This section confirms the safety of our product. As we've communicated with experts from different fields, the safety of our product can be optimized and verified in terms of every aspect.

Implementation

This section explains the application of our products to specific stakeholders. With the support of other experts, we received lots of suggestions and made some improvements in order to better meet the needs of stakeholders.

Business

This section allows us to analyze and improve the feasibility of project entrepreneurship from a more professional business perspective. It also indicates the economical impacts bring by our project.

Laws and regulations

This section can ensure that our project complies with local laws and regulations. This will help our project better enter the real world. Here, the patentability and legitimacy of our products will be demonstrated.

Our journey began when our team members were troubled by mosquitoes, and the mosquito control products currently available on the market did not meet their needs. After literature research, it was found that mosquito borne diseases pose a threat to people around the world. Through communication with disease control experts and citizens, we have gained an understanding of the harmfulness of mosquitoes, the necessity of mosquito control, and the demand for new mosquito killing products, which has helped us identify potential end users. Based on the needs of potential target customers, we consulted with biosafety and technology experts, and our solution gradually took shape. At the same time, in order to expand the influence of our project, we have raised people's awareness of mosquito control through educational classes and lectures, further expanding our target customer group. We also consulted experts in the business and legal regulatory industries to lay the foundation for Moskilla's entry into the market.

In summary, our human practices journey mainly consists of four stages, namely identifying issues, formulating our solution, expanding project impact(education), and commercialization(IHP sth about business).

You can learn about the events that occur during our journey through the rectangular text box. Each outreach has flag markers with different colors in the top right corner to indicate for which event category. If you are interested in our reflection process at an event, please click the READ MORE button to learn about our analysis of applying the AREA framework. You will understand our purpose, gains, reflections, and outlook.

∙ Our Timeline

In summer, along with the rising temperature, people's daily lives and sleep are troubled by active mosquitoes, and the demand for mosquito repellent products is also increasing. However, people rarely realize that mosquitoes are the deadliest animals in the world through transmitting diseases.

Nowadays, mosquitoes have highly affected the human race through transmitting diseases. According to researches, dengue fever, transmitted by mosquitoes, is prevalent all over the world: about 400 million people are infected and 21,000 deaths are caused by it annually (CDC, 2018).

Figure 2. Global distribution of Dengue Fever in 2018

However, mosquitoes transmit not only dengue, but also malaria and many others. An estimated 249 million cases of malaria occurred in 85 malaria-endemic countries in 2022 (CDC, 2024). Of the 249 million, approximately 608,000 died of malaria. Taking all mosquito-borne diseases into account, mosquito-borne diseases contribute to 17% deaths among all infectious diseases (WHO, 2020).

Furthermore, mosquito-borne diseases also lead to massive economical losses: the estimated cost of medical care for the West Nile virus diseases, transmitted by mosquitos, are over $56 million dollars per year in the U.S. alone (West Baton Rouge Museum, 2019). These statistics demonstrates that the danger of mosquitoes’ prevalence is extremely obvious and needs to be dealt with.

Figure 3. Significant damages caused by mosquitoes

∙ Local Issue

China has always been heavily impacted by mosquito-borne diseases. China was once strongly affected by malaria, reporting over 24 million cases in the early 1970s (WHO, 2024). In 2010, the government launched the National Malaria Elimination Programme, effectively mitigated the problem. Starting from 2017, China reported zero malaria cases, making itself a malaria-free country recognized by the WHO.

However, while malaria is eliminated, dengue remains its significance in China. In 2023 alone, over 5,000 cases of Dengue Fever were reported in China (National Disease Control and Prevention Administration, 2023). In the post-COVID era, the transmission of various mosquito-borne diseases, especially Dengue Fever, is likely to rise in China, posing new challenges in disease control. Novel prevention and control methods need to be developed to improve the current situation.

Figure 4. Dengue prevalance in China 2005-2020

∙ Significance of mosquitoes control

Mosquito-borne diseases pose serious threats to health of the humanity. However, many mosquito-borne diseases are preventable. Through strategies eliminating or controlling the mosquito population and the contact between humans and mosquitos, the spread of these infectious diseases could be restricted. Therefore, the authorities are now endorsing mosquito control, encouraging people to actively kill mosquitoes. For instance, the Centers of Disease Control and Prevention (CDC) encourages individuals and community directors to kill mosquitoes, and provides guidance for mosquito control.

Figure 5. CDC's guidance for mosquito control

∙ HMBPP attracts blood seeking mosquitoes

(E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP), known as an isoprenoid precursor, has been revealed also a phagostimulant for blood-feeding mosquitoes. It was first found that the parasite Plasmodium falciparum produces HMBPP during malaria infection to indirectly increase the release of carbon dioxide, aldehydes, and monoterpenes in red blood cells. It enhances attraction and feeding behaviors of mosquito vectors. (Emami et al., 2017) Based on its phagostimulatory effect, the addition of 10 uM HMBPP to plant-based solution in combination with chemical toxins has been shown to have potential as an environmentally friendly ATSB for mosquito control (Viktoria E. S. et al., 2021).

This inspired us to use HMBPP to attract those blood-feeding mosquitoes. However, HMBPP can not yet be chemically synthesized. Therefore, we have decided to choose E. coli as the chasis cell producing HMBPP, as it has the same natural MEP pathway as Plasmodium. Also, E. coli is a type strain which can be easily engineered to increase the yield of HMBPP. (Zhou et al., 2017)

Figure 6. Compared to sugar baits, blood-seeking mosquitoes prefer HMBPP

∙ RNAi yeast for mosquito control

RNA interference (RNAi) is a post-transcriptional gene silencing mechanism that operates through the sequence-specific degradation of target mRNA transcripts, mediated by small RNA molecules (sRNA). Given the increase of reported chemical insecticide resistance and the rising concern for negative effects of broad spectrum toxic mosquitocides on non-target organisms, there is an urgent need for the development of novel, environmentally-friendly mosquitoicide. To this end, RNAi approaches are pursued as a promising strategy for mosquito control.

Whereas, both application in ATSBs and large-scale production require the stability and efficient uptake by mosquitoes. In this regard, dried and inactivated yeast short hairpin RNA (shRNA) pellets has been identified as an optimal approach to deliver shRNA mosquitocide. This method has demonstrated the highest efficacy in inducing mortality across various mosquito species, while simultaneously achieving the silence of targeted genes. (Molly D. S., 2019)

For the purpose of gene silencing, genes that are pivotal to mosquito fecundity, behavioral patterns, survival, and vector competence are prioritized. In our study, we have selected the 5-HT1 receptor (5-HTR1) gene, which encodes serotonin receptors present on neuronal and other cellular membranes, and the Rbfox1 gene, encoding an evolutionarily conserved RNA-binding protein. The previous researches indicate that silencing either the 5-HT1 receptor or Rbfox1 leads to the death of mosquitoes, with negligible impacts on non-target organisms. (Keshava et al., 2021; Keshava et al., 2023)

Figure 7. RNAi mechanism activated in the body of mosquitoes consuming targeted shRNAs

Now, let us introduce a novel solution, Moskilla, which is an optimized ATSB containing two main components: HMBPP addition sugar bait to attract mosquitoes and targeted shRNA to kill mosquitoes. Our objective is to develop an eco-friendly, safe, effective, and highly specific method for mosquito control. Moskilla is mainly composed of the following 2 components.

HMBPP addition sugar bait: Except traditional sugar bait, HMBPP producing E. coli is also added. The presence of HMBPP and sugar in Moskilla can optimize its attractiveness to both sugar- and blood-feeding mosquitoes. To reduce the risk of E. coli leakage, the engineered E. coli has been constructed to a nutrient-deficient strain, thereby limiting its survival outside our controlled application context.

shRNA mosquitocide: shRNA mosquitocide of Moskilla is encapsulated within dried, inactivated yeast cell pellets that express the targeted shRNA sequences. This formulation is designed to initiate the RNA interference (RNAi) mechanism upon ingestion, effectively silencing vital mosquito survival genes, such as 5-HTR1 and Rbfox1. Our shRNA sequences are rigorously designed to minimize off-target effects, thereby significantly reducing the toxicity to other non-targeted organisms compared to conventional chemical mosquitocides. This innovation renders Moskilla a more sustainable, safe, and highly targeted approach to mosquito control.

Figure 8. Composition of Moskilla