Describe the goal of your project: what is your engineered organism (or other synthetic biology product, system, or tool) supposed to do? Please include specific technical details and names of important parts. Our project aims to modulate the rapid translation of mRNA-based vaccines with the international to reduce the severity of symptoms experienced. We plan to do this by regulating the delivery of mRNA in mammalian cells through the use of mRNA folding with the software package to predict the ideal insertion of a translation regulating structure known as an iron-responsive element and its associated iron-binding protein.
Which whole organisms, including viruses and cell lines, are you planning to use (or using) in your project? CHO cells, E. coli K-12, and E. coli DH5alpha
What risks could these organisms pose to you or your colleagues in the laboratory, or to your community or the environment if they escape the lab? Transgenic e. Coli and mammalian cells pose risk to the environment if not handled and disposed properly. All contaminated equipment will be autoclaved to ensure safety. Benchtops will be sterilized after use with isopropanol. Toxic reagents will be disposed of properly according to standard lab safety procedures.
What organisms are you using as chasses in your project? Escherichia coli: E. coli, K-12, E. coli DH5alpha Others: CHO cell
What risks could your chassis pose to you or your colleagues in the laboratory, or to your community or the environment if they escape the lab? E. coli poses risk to the environment if not handled and disposed of properly. All contaminated equipment will be autoclaved to ensure safety. Benchtops will be sterilized after use with isopropanol.
As part of your project, are you planning to make/have made new parts or substantively changed existing parts in the Registry? Yes
Which chemicals are you using in your project that might be hazardous? Heavy metals, Carcinogens, Mutagens, Acids, and corrosive chemicals.
What experiments will you do with your organisms and parts? E. coli DH5apha transformation. CHO transfection with our plasmid designs. We will be observing changes in fluorescence with a microplate reader.
What risks could arise from these experiments? A potential hazard that may arise from our experiments is exposure to specific mutagenic compounds such as ethidium bromide and mammalian cell excretions. There is also the concern of exposure of our own mRNA designs if ingested and potentially can translate in resulting in the innate immune response.
Are you collecting any data about people, such as their opinions, quotations, health information, gender, behavior, attitudes, or concerns? Yes
Imagine that your project was fully developed into a real product that real people could use. How would people use it? Only in the lab or to be used in the human body, or in the food
What safety, security or ethical risks would be involved with such a use? Developing a way to improve vaccines by decreasing adverse side effects would pose safety risks down the line in clinical tests, as the promise of fewer adverse side effects could mislead people, and potentially cause those who are at high risk, like immunocompromised individuals or elderly individuals, to get vaccinated in the future without still considering the side effects.
If you were permitted, would the continued development of your project require environmental release (as described in our Do Not Release policy)? No
If your project were fully developed, could any of your engineered organisms or parts spread autonomously in the environment? no
How will experts overseeing your project help to manage any of the risks you identified in this form? Biological Use Authorization officers are responsible for the safety and security of biology labs. Work with biological agents or recombinant DNA requires a Biological Use Authorization (BUA) from the UC Davis Institutional Biosafety Committee (IBC).
What rules or guidance cover your work which could help to manage any of the risks you identified in Part 2 of this form (in particular Question 11)? https://asm.org/getattachment/3cleb38c-84d7-472f-aa9b-5d695985df21/ASM-Biosafety-Guidelines.pdf
Have your team members received any safety training? Yes, we have already received safety training.
Please select the topics that you learned about (or will learn about) in your safety training. Lab access and rules, responsible individuals, differences between biosafety levels, biosafety equipment, good microbial technique, disinfection and sterilization, emergency procedures, transport rules, physical biosecurity, personnel biosecurity, dual-use and experiments of concern, data biosecurity/cyberbiosecurity, chemicals, fire and electrical safety.
Which work areas do you use / are you using to handle biological materials? Open bench, biosafety cabinet, chemical fume hood, separate cell culturing space.
What is the biosafety level of your work space? Level 1
What other risk management tools will cover your work? Accident reporting, personal protective equipment, an inventory control system, access controls, medical surveillance, waste management system, special procedures or protocols that address safety or security.
How will the rules, training, containment and other procedures and practices help to manage any of the risks you identified in Part 2 of this form (in particular Question 10)? All contaminated equipment will be autoclaved to ensure safety. Benchtops will be sterilized after use with isopropanol. Toxic reagents will be disposed of properly according to standard lab safety procedures. Dr. Albeck will be supervising us throughout the experiments.
Are you planning to release any organism or product derived from your project? No
Are you planning to use (or using) any animals (including insects and invertebrates) not on the Whitelist? No
Are you planning to use (or using) any vertebrates (e.g. rats, mice, guinea pigs, hamsters) or higher order invertebrates (e.g. cuttlefish, octopus, squid, lobster)? No
Are you planning to use (or using) any parts not on the Whitelist? No
Are you planning to do (or doing) any of the activities not on the Whitelist? No
Are you planning to use (or using) any parts or organisms obtained from anywhere other than a trusted commercial or institutional supplier? No
Are you planning to use or test any part of your project, or products produced from it, on humans (including members of the team)? No
What else can you tell us about any risks associated with your project, how you are managing them, or your compliance with iGEM’s safety and security rules and policies? n/a