- The incidence of Dengue has grown dramatically around the world in recent decades. According to WHO or the World Health Organization, the cases reported increased from 505430 cases in 2000 to 5.2 million in 2019. Furthermore, social and environmental factors including unplanned urbanization, water storage practices, rapid population growth, and etc. In addition, Dengue is a highly transmissive disease, where every infected mosquito will stay viremic for its whole life and symptoms of infected people get more deadly if they are infected again. NS3-Hel and NS3-ZIKA are important proteins in Dengue or Flavivirus.[1] Helicase plays an important role in viral replication and is a very promising specific antiviral inhibitor launch target. Since there are still no way to cure Dengue, and the infected cases keep increasing dramatically, we want to invent medicine that could suppress the function of the helicase. [2]

- In our experiment, we first use PCR cloning to rapidly replicate the two DNA fragments: NS3-Hel and NS3-ZIKA. Then we use gel electrophoresis to separate DNA according to their molecular sizes. To isolate these DNA from the agarose gel, we use DNA Gel Extraction kit. Now we have the NS3-Hel and NS3-ZIKA DNA ready to connect with the plasmid. In order to create a space for the two bacteria, we want to use enzyme digestion. We finish construct the plasmid containing the two bacteria. Next step is to be able to express the gene, where we use heat shock induced transformation. We do a PCR testing to see if the DNA is successfully transferred into the BL21(DE3). Then we will do a protein electrophoresis to test if the protein is successfully expressed. Finally we want to test the activity of helicase by checking whether it has fluorescence or not. [2]

- Our goal is to invent medicine that could inhibit the helicase activity, which would stop the Dengue from replicating. We aim to solve the stalemate where proper therapeutic methods to treat Dengue are lacking. We want to develop a drug screening kit that is fast and efficient to test if the substance contains the ability to inhibit the helicase activity. [3]

[1]. World Health Organization. (2023). Dengue and severe dengue. World Health Organization. https://www.who.int/news-room/fact-sheets/detail/dengue-and-severe-dengue

[2]. Belon, C. A., & Frick, D. N. (2008). Monitoring helicase activity with molecular beacons. BioTechniques, 45(4), 433-442. https://doi.org/10.2144/000112834

[3]. Blaney, J. E., Jr, Hanson, C. T., Firestone, C. Y., Hanley, K. A., Murphy, B. R., & Whitehead, S. S. (2004). Genetically modified, live attenuated dengue virus type 3 vaccine candidates. The American journal of tropical medicine and hygiene, 71(6), 811-821.