In the modern era, responsible and safe research is a top priority for both the subjects it applies to and the scientists that carry it out. Even though our team did not have access to a lab over the course of this summer, we were still required to complete safety training. All researchers at Stony Brook University must complete an array of assesments and modules demonstrating knowledge of lab safety procedures, waste disposal, and procedures that must take place in the event of an accident. Our lab at the Centers for Molecular Medicine has a Biosafety Level 1 rating, meaning it requires us students to utilize safety protocols such as decontamination of all materials prior to disposal via autoclave, use of Personal Protective Equipment (PPE) such as nitrile gloves, ANSI Z87.1 safety goggles and long lab coats, and prohibition of all food, drinks, and smoking materials inside of and around the lab.
Due to the safety measures introduced after the onset of COVID-19, Stony Brook iGEM has not been granted access to any labs. Instead, our team is worked from home and put a greater emphasis on other aspects of the project.
We hope to introduce an optogenetic transcription control system into Nicotiana benthamiana leaves which will prevent plant development upon exposure to UV-B light. This optogenetic transcription control system will utilize ULTRAVIOLET RESPONSE LOCUS 8 (UVR8) and its binding partner CONSTITUTIVE PHOTOMORPHOGENIC1 (COP1) to control the transcription of synthetic trans-acting small interfering RNAs (syn-tasiRNAs) upon exposure to UV-B light (~311 nm). These syn-tasiRNAs will be transported to the shoot apical meristem (SAM) via phloem, where they will bind to WUSCHEL (WUS) mRNAs, forming an RNA-induced Silencing Complex (RISC). The resulting knockdown of the WUS gene will hopefully interfere with the CLAVATA-WUSCHEL signaling pathway, causing stem cells within the SAM to differentiate. This stem cell depletion would ultimately prevent growth of the whole plant.
Our constructs and research design center around the use of Nicotiana benthamiana but, since all of our work will be done solely online, we will actually not be using any organisms.
We hope to incorporate our optogenetic killswitch in a variety of transgenic plants. Given that the core signaling component of the CLV-WUS pathway is relatively conserved in a diversity of crops such as tomato (Solanum lycopersicum), maize (Zea mays), and rice (Oryza sativa), preliminary implementation will include the integration of our constructs on these plants. For a more detailed description of how we hope to complete this, feel free to visit our Proposed Implementation page.
Safety, Security, and Ethics
Our experiments are closely tied to genetically modified organisms (GMOs). Therefore, there are some ethical concerns that must be addressed. Weale (2010) explains a total of 5 ethical concerns that have risen about genetically modified crops: potential harm to humans, potential damage to the environment, potential negative impact on traditional farming methods, potential of corporate dominance, and the apparent "unnatural" stigma of technology and genetic engineering.
Our advisor Dr. John Peter Gergen, head of Undergraduate Biology at Stony Brook University, oversaw our project to help us manage any risks if they are to arise. Dr. Gergen's work revolves around genetics, currently with a primary focus on the Runt transcription factor. Our project doid not have major safety or security concerns, as we were not working with toxins or viruses. We also did not work in a lab or conduct any experiments.
In preparation for wetlab work, all members of the Stony Brook iGEM team must complete a Collaborative institutional Training initiative (CITI) on responsible conduct of research. Stony Brook safety courses are required as well to teach students waste disposal and different hazards. These courses include dealing with hazardous wastes (ENV 001), chemical hazards (ELS 002), biological hazards (ELS 003), and regulated medical waste (ENV 005). More information on these courses can be found here.
Weale A. (2010). Ethical arguments relevant to the use of GM crops. New biotechnology, 27(5), 582–587. https://doi.org/10.1016/j.nbt.2010.08.013