Project Description
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As first time iGEMers, we did not have a clear idea of what our project focus should be when we recruited our members. After having a meeting together, we decided to work on a topic that would be helpful inside of our school, especially among the secondary community. So, we gathered results from an online survey filled out by our secondary students (G7-G12) which asked for common problems that they encounter from a student perspective. The most frequent reply we observed was sleeping problems, with some other replies being also very relevant.
Our team then had a meeting, and we decided to settle down and tackle the sleep problems of our schoolmates. We then conducted a second online survey which aimed to learn more about the sleeping habits of our students and to obtain more detailed reasons for their sleep problems, from the results we concluded that students have no issues during their sleep itself, but rather have issues getting to sleep and being unable to wake up refreshed and energized in the mornings. After a few other meetings, in which we brainstormed about what the most suitable solution to our community’s problem could be, aromatherapy rose above other methods to help our peers with their sleep problems.
About Aromatherapy Aromatherapy is a therapeutic practice deeply rooted in ancient traditions, which uses natural aromatic compounds, known as essential oils, to promote physical and psychological well-being [1]. Aromatherapy in modern society is used as alternative and subsidiary medicine, serving as a secondary treatment that is paired with other standard treatments, with the intention to boost the effects of the main treatment [1]. In China, there is a long history of aromatherapy practice, with the earliest record found being about 2500 BC ago, during the reign of Huang Ti, the Yellow Emperor [2]. In today’s world of aromatherapy, China is one of the most prolific producers of essential oils and has contributed many valuable historical plant medicine text records [2]. The particular scent of many plants, including citruses and laurels, are given by the terpenes produced in their metabolic pathways, making them very appealing for the production of essential oils as these flowers are usually rich in terpene content [3][4]. Essential oils are made by extracting the chemical constituent volatiles from the original plant using steam distillation [3], to further separate the terpene molecule content from the essential oil, methods such as using very nonpolar organic solvents or breaking the plant cells to release the terpene compound are often the choice of producers [4]. When inhaled, the terpene molecules travel from the olfactory nerves directly to the brain and affects the amygdala, the emotional center of the brain. Research showed that aromatherapy can be used to improve sleep quality [5] and boosting focus and awareness [6].
Our Approach To help our students to fall asleep easier and to wake up refreshed we aim to produce recombinant yeast that is able to synthesize menthol and vanillin as our final goal, providing mint and vanilla scents respectively. Mint scent would improve focus and refresh our students due to its cooling sensation when inhaled [7] and vanilla scent would act as an agent to improve sleep quality [8]. As can be seen in Fig. 1, the primary biosynthetic step of menthol is limonene, we will thus begin by synthesizing and expressing the limonene terpene synthase genes geranyl diphosphate synthase (GPPS) and limonene synthase (LimS). Similarly, we will synthesize the vanillin terpene synthase gene fcs that converts ferulic acid to feruloyl CoA and ech gene that converts feruloyl CoA into vanillin as illustrated in Fig. 2. This will first be done in E. coli for testing, if successful we will then clone the target gene sequaences into yeast cells.
When considering how students can switch between the relaxing vanilla scent at night before they go to bed and the refreshing menthol scent in the morning when they wake up, we decided to design the plasmid so that the synthesis of limonene will be activated by red light, using the PhyB-PIF system from plants [11][12], and the synthesis of vanillin will be activated by blue light, using the pDawn system that express the genes under a blue light-responsive promoter [13]. If we can successfully synthesize vanillin and limonene, we aim to continue with the metabolic pathway to try and synthesis limonene to menthol. For ease of use we are also designing a device, as can be seen in Fig.3, that can be set to on a timer to expose our transgenic yeast to blue or red light as needed. This way students can set the timer to either expose the yeast to red light in the morning, so that the yeast will produce the fresh menthol scent in the morning when they need to wake up, or to expose the yeast to blue light at night before they go to bed, so that the yeast can then produce the calming vanilla scent to help them fall asleep.
Refrence
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-[1]: Farrar AJ, Farrar FC. Clinical Aromatherapy. Nurs Clin North Am. 2020 Dec;55(4):489-504. doi: 10.1016/j.cnur.2020.06.015. Epub 2020 Sep 28. PMID: 33131627; PMCID: PMC7520654.
-[2]: Baser, K. H. C., & Buchbauer, G. (2009). Handbook of essential oils: science, technology, and applications. CRC press.
-[3]: Aziz, Zarith & Ahmad, Akil & Mohd-Setapar, Siti & Karakucuk, Alptug & Azim, Muhammad & Lokhat, David & Kamal, Mohammad & Ashraf, Ghulam. (2018). Essential Oils: Extraction Techniques, Pharmaceutical And Therapeutic Potential - A Review. Current Drug Metabolism. 19. 10.2174/1389200219666180723144850.
-[4]: Elshafie HS, Camele I. An Overview of the Biological Effects of Some Mediterranean Essential Oils on Human Health. Biomed Res Int. 2017;2017:9268468. doi: 10.1155/2017/9268468. Epub 2017 Nov 5. PMID: 29230418; PMCID: PMC5694587.
-[5]: Salamung, N., & Elmiyanti, N. K. (2023). Effect of Aromatherapy on Sleep Quality: A Systematic Review. International Journal of Nursing and Health Services (IJNHS), 6(5), 292–302. https://doi.org/10.35654/ijnhs.v6i5.728
-[6]: Diego, M. A., Jones, N. A., Field, T., Hernandez-reif, M., Schanberg, S., Kuhn, C., … Galamaga, R. (1998). Aromatherapy Positively Affects Mood, Eeg Patterns of Alertness and Math Computations. International Journal of Neuroscience, 96(3–4), 217–224. https://doi.org/10.3109/00207459808986469
-[7]: Eccles R. Menthol and related cooling compounds. J Pharm Pharmacol. 1994 Aug;46(8):618-30. doi: 10.1111/j.2042-7158.1994.tb03871.x. PMID: 7529306.
-[8]: AL-SANDOOK, T., KHALIL, B., & TAWFIK, N. (2020). Vanillin soothing effect in treating certain oral disease entities. Al-Rafidain Dental Journal, 1(2), 143-147. doi: 10.33899/rden.2020.166034
-[9]: Croteau RB, Davis EM, Ringer KL, Wildung MR. (-)-Menthol biosynthesis and molecular genetics. Naturwissenschaften. 2005 Dec;92(12):562-77. doi: 10.1007/s00114-005-0055-0. PMID: 16292524.
-[10]: Zhang X, He Y, Wu Z, Liu G, Tao Y, Jin JM, Chen W, Tang SY. Whole-Cell Biosensors Aid Exploration of Vanillin Transmembrane Transport. J Agric Food Chem. 2021 Mar 17;69(10):3114-3123. doi: 10.1021/acs.jafc.0c07886. Epub 2021 Mar 5. PMID: 33666081.
-[11]: Hughes RM, Bolger S, Tapadia H, Tucker CL. Light-mediated control of DNA transcription in yeast. Methods. 2012 Dec;58(4):385-91. doi: 10.1016/j.ymeth.2012.08.004. Epub 2012 Aug 15. PMID: 22922268; PMCID: PMC3526682.
-[12]: Tabor JJ, Levskaya A, Voigt CA. Multichromatic control of gene expression in Escherichia coli. J Mol Biol. 2011 Jan 14;405(2):315-24. doi: 10.1016/j.jmb.2010.10.038. Epub 2010 Oct 28. PMID: 21035461; PMCID: PMC3053042.
-[13]: Robert Ohlendorf, Roee R. Vidavski, Avigdor Eldar, Keith Moffat, Andreas Möglich, From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression, Journal of Molecular Biology, Volume 416, Issue 4, 2012, Pages 534-542, ISSN 0022-2836, https://doi.org/10.1016/j.jmb.2012.01.001. (https://www.sciencedirect.com/science/article/pii/S0022283612000113)