Parts

      In our experiment, we constructed four vectors, including two DlGAD overexpression vectors (BBa_K4113022 and BBa_K4113025) and two gene editing vectors to knock down DlCaM. Among them, CaM knockdown treatment can significantly increase the content of GABA in Longan callus. Our team has uploaded the relevant components, a total of 15, to provide some reference for future iGEM teams interested in the direction of GABA synthesis. You can see the details in the parts section!

Mathematical Modeling

      Through extensive literature review, we know that the simultaneous stress of high light intensity and high temperature leads to a unique metabolic response in plants, and some scholars have found that GABA content is increased under both treatments after rigorous identification [1].
      However, due to the epidemic situation, the content of our experiment was limited. Only 40℃ high temperature stress treatment and 32umol·m-2·s-2 blue light treatment with a specific single parameter were carried out on Longan callus, without exploring the blue light treatment of other temperature and parameters, as well as various complex combination treatments before. We hope to continue the exploration and verification of this part by establishing a reasonable mathematical model for the experiment.
      Our project aims to establish a GABA-producing longan cell line. In order to simulate the work of this cell line, we conducted experiments in suspension culture to simulate GABA production in small light-sheltered fermenters. Part of our modeling implementation is to experiment difficult to carry out factory standard models under ideal conditions for scaling up cultivation. Our cell line culture takes weeks as the time unit, and carries on the renewal. Through the batch measurement of GABA content of suspended materials within a week, the basic model of GABA content change is simulated. Based on this model, we modeled again to find out the most suitable time for GABA extraction as well as the corresponding incubation days and batch number, so as to propose a reliable production model for large-scale factory production. Provide the basis for standardized production. In addition, we also modeled the location of Longan GABA plant cell factory, and found that Putian City in Fujian Province was the best location, which could meet the basic growth needs of Longan, which provided certain suggestions for the location of Longan cell factory.

Human Practice

      Recent years, a considerable group of people is troubled by anxiety, violent mood swings and insomnia. These phenomena could be draw by our increasing speed of social development, which call for solution and are not mitigated yet. Fortunately, although these problems are ignored rather than addressed, we have been always finding ways to prevent everyone from falling into the trap of mood. To cope with it, we have carried out both experiment and human practice to assist all with psychological problems. Contributions are mainly divided into two parts as follows.
         To understand the current situation of anxiety and insomnia, except for handing out questionnaires, we went deeply into kindergarten, families and governments to investigate and interview.
      For the education part, we succeeded in scientific popularization in schools, organizations and Yuxi Town, a countryside in Fujian Province. Moreover, our team have cooperated with several other groups such as Yahao Media Company, BUCT-China and other iGEM teams. The sound results of education and reflection from our audiences are achieved in various ways. Apart from the traditional education patterns, we also set up a official account to spread our practice and share knowledge universally.       Generally, in higher plants, the content of GABA in tissues is generally within 0.3-32.5μmol/g. Low GABA content and relatively difficult extraction limit its production and application. Our team used gene editing technology to obtain high-GABA cell lines, while expanding cultures through fermenters to build a high-yielding GABA cell factory. Pharmaceutical companies could use our ideas to extract GABA for new drugs, which could play an important role in alleviating insomnia, stress and so on.
      After communicating and learning with beverage companies, our team combined with the advantages of the project to get inspiration and integrate the extracted GABA into daily life. So far, we've created a high-GABA beverage that beverage companies can work with us to mass-produce as a sweeter solution to insomnia.

Reference
[1] Balfagón, Damián et al. “γ-Aminobutyric acid plays a key role in plant acclimation to a combination of high light and heat stress.” Plant physiology vol. 188,4 (2022): 2026-2038. doi:10.1093/plphys/kiac010