In Brazil, ANVISA allows the use of medicinal cannabidiol while simultaneously prohibiting the domestic cultivation of Cannabis sativa for its extraction. Consequently, the regulation encounters ethical dilemmas, notably concerning the responsibilities of healthcare professionals in prescribing and monitoring its usage. Furthermore, society taboos and biases linked to the plant's association with recreational Cannabis use are prevalent (Longarai, 2023).
In response to these challenges, our project presents a solution: we aim to enhance accessibility to this substance through a synthetic biology approach. This innovative method avoids the need for plant cultivation and aims to significantly reduce ethical dilemmas and societal taboos surrounding its production and use. Therefore, our proposal seeks to democratize access to this substance, especially for individuals afflicted with diseases that require its therapeutic benefits, with a particular focus on epilepsy.
Our project is focused on synthesizing self-cleaving proteins using the T2A sequence, an advanced genetic engineering technique that merges segments of diverse genes into a single fusion gene, which is transcribed into a single mRNA and translated into several protein molecules. This methodology enables wide-ranging applications such as tailoring therapeutic proteins to individual needs and facilitating the investigation of biological processes (Wang & Marchisio, 2021).
In order to express these enzymes in Saccharomyces cerevisiae yeast, we built biological circuits, analyzing and organizing their components, such as promoters and terminator, plasmids and for those enzymes that needed, setting the components in an expression cassette. So, we proposed circuit names, to express their respective proteins.
Figure 1: Circuit with PT4 and CBDAS.
Source: 2023, Authors.
Figure 2: Circuit with HCS, OS and OAC.
Source: 2023, Authors.
We also made some analysis of catalytic site and molecular docking for the proteins modeled in our project. These analyses could assist the studies of the enzymatic behavior for reactions in the pathway we studied. Furthermore, these analyses contributed to the general knowledge around these enzymes, by elucidating possible binding and catalytic sites that were not yet described in literature (see Model page, Phylogenetic Analysis at Molecular Docking section).
Furthermore, our team contributed to the iGEM community by adding new parts to the registry. The parts added not only are ready-to-use devices, but also contain highly interchangeable parts and are applicable in several eukaryotic expression systems, which the entirety of future iGEM teams and the scientific community may explore. We added 2 devices, 2 composites, 1 improved part, some slightly modified parts and several primer sets designed for our cloning strategy into the pRS vector series, but the parts can be easily inserted into other plasmids by designing new, adequate primers. Check out all of our designed parts either on this Wiki’s Parts page , or on our team’s pages on the Registry!
In addition to our hard work with modeling and building biological circuits, we also accepted the challenge of thinking outside the box in order to make a useful contribution for future iGEM teams. To do so, we spared no effort in the material left by our Human Practices team.
As this competition promotes a collaborative environment in which teams can exchange information and use discoveries made by other teams in previous years to support research and new projects, we feel thrilled to be able to participate and bring new resources and data that can be somehow repurposed afterwards.
To be honest, writing this text is even a bit emotional for us. CBDynamics is a project that has been "on the back burner" for almost five years. As we've said in some sections of this wiki, it's a project that tackles a controversial issue in our region. This year, some factors have been essential in giving us the courage to face up to the dream of working on this project. As we read the wiki of several past teams, we found ourselves really inspired and excited about making it work.
Taking into account the above, as well as the desire to inspire teams in the future in the way we were inspired, we dedicated a lot of our Human Practices session to showcasing refined research on the impact of our project in the different spheres it can encompass: Academia, Local Community, Industry, Healthcare and impact on Government and Public Organizations.
Taking into account the above, as well as the desire to inspire teams in the future in the way we were inspired, we dedicated a lot of our Human Practices session to showcasing refined research on the impact of our project in the different spheres it can encompass: Academia, Local Community, Industry, Healthcare and impact on Government and Public Organizations.
Of course, in addition to refined research, we also consulted various opinions. It's important to note, however, that opinions are always very much related to the cultural environment, and can change greatly from neighborhood to neighborhood, city to city, state to state and country to country. The data we collected can be useful in broadening the range of people we listen to. After all, when talking about real people trying to solve real problems, we can't help but listen to each community and its demands.
A lot of people are not so open to the subject, not because they have well-conceived opinions on the matter, but out of sheer ignorance. When we sat down to talk about it with many people, the feedback we received was actually very positive, as if a new horizon had been opened up for them. Unfortunately, we have no way of talking to all the 80,000 people who live in our city, let alone the 214 million people who live in Brazil, or the 7.8 billion people who inhabit this planet.
For those of you reading our wiki, we swear that if there was a way, we'd be crazy enough to do it. But the 22 engineering students who make up this team haven't managed to solve this problem.
These students (us) did, though, manage to find what we call the advent of change. The solution even seems a little obvious (but make no mistake, "obvious" and "easy" are not synonymous). What better way to reach the population and bring about change than to focus on children?
With that in mind, our main contributions to the HP field are related to the production of high-quality classroom materials focusing on an introduction to synthetic biology, biological circuits and the medicinal benefits of CBD. This material can be useful for teams working with cannabidiol in the future, exploring alternative treatments or trying to combat pseudoscience in general. There is no shortage of controversial topics in science. There is no shortage of topics that are taboo for a large part of the population. It's not just our project that is controversial, but many other topics too. It is not at all effective to explain to a classroom only about the difference between the use of cannabidiol as a treatment for illnesses and the recreational use associated with Cannabis consumption or smoking. This discussion is valid and necessary, but it is also necessary from an early age to explain how solutions to various types of problems are tested by science and the ethical discussions behind it all. In this way, news that is often wrongly propagated through ignorance will have less and less place in the world.
Future teams can also benefit from our data collection, whether it's to compare it with other scenarios around the world or even to compare the evolution of popular opinion on such a controversial topic over the years.
We are also very positive that our way of approaching this issue has been very fruitful and could be replicated by other teams who, like us, tackle projects that touch on such polemic topics. If we can also leave a piece of advice, we would like to say that we realized that approaching the topic with the population in a light-hearted way and bringing a perspective that mixes visions and inserts the subject into daily topics is the best way not to cause the general population strangeness to the project.
Longarai, G. (2023). Canabidiol medicinal: direito à saúde no Brasil e o papel do judiciário em meio a conflitos legais, morais, tabus e preconceitos. Porto Alegre, Brasil. Retrieved from https://repositorio.animaeducacao.com.br/bitstream/ANIMA/33791/1/ARTIGO%20GEORGE%20SOLON%20LONGARAI%20.pdf
Wang, X., & Marchisio, M. A. (2021). Synthetic polycistronic sequences in eukaryotes. Synthetic and Systems Biotechnology, 6(4), 254–261. https://doi.org/10.1016/j.synbio.2021.09.003
Bonde, J.S., Bulow, L. (2013) Chimeric Genes, Proteins, Brenner’s Encyclopedia of Genetics, Second Edition, Elsevier.
Biological circuits designed with SnapGene software (www.snapgene.com).