Why saftey is important
iGEM has 2 major guidelines for laboratory work during a project:
1. No organisms of riskgroups 3 or 4 should be used.
2. No genetically modified organisms should be released outside the lab.
The first guidline does not apply since all our parts and organisms are classified as riskgroup 1 at the highest. However, since we modify a species (C. reinhardtii) which can be applied to the second guideline. This rule becomes especially crucial if the organism are classified as a GMO.
The question therefore follows; should our projcet be considered as GMO? Yes we do have a genetically modified eukaryotic organism, however despite it being genetically modifed, by defintion it’s not to be considered as GMO since we have been using CRISPR-Cas9 system to cut a gene without inserting a new DNA. The FDA (citation), aswell as Jordbruksverket (citation) considers the use of CRISPR-Cas9 not to be GMO if you use the system only to cut a gene. Despite this we are not planning on releasing it outside the lab except for submitting the parts into the registry.Safe Project Design
At LiU iGEM our main concern has always been safety. The decisions involving project design has therefore carefully been analyzed to minimize the amount of potential risks. The chassis we are using are a non-pathogenic XL-1 blue E.coli strain and all the parts included are unharmful to humans and other organisms. The main project challenge was to ensure that our genetically modified C.reinhardtii would not pose any threat to other wildtype alga. That’s being satisfied by the fact that Cas9 and the sgRNa will severely weaken the modified C. reinhardtii to the point that it will not be able to reproduce. This is especially true since our goal is to eventually modify the chloroplast, which, when modfied will severly weaken C.reinhardtii to grow outside in the wild. To assure that no modified C.reinhardtii would escape into the wild this system will be activated by light, so that ordinary sunlight would be sufficient to shut down the organism.
Safe Lab Work
Saftey is paramount when working in the laboratory, and this project is no expection to this. Before each lab a risk assesment was written down. Since we were using riskgroup 1 organisms we were working on a lab bench and always wearing lab coats, saftey glasses and latex/nitrile gloves. The algae were treated as though they are riskgroup 1 organsims. The waste of both bacteria and algae were collected in separate waste containers and then sterilized through autoclavation. This was also true for all the solid waste that had touched alage or bacteria. To ensure no mutated bacteria or algae got outside the lab, each laborant sterilized their hands aswell as the lab bench they were working with. For chemicals that were smelly (i.e acetic acid), or poisionous (i.e hygromycin) or any other types of hazards was handled under a fumehood with nitrile gloves and saftey glasses with outmost care when handling them. The most dangerous chemical that we worked with, ethidum bromide, were handled with extreme care, that means always double nitrle gloves and under a fume hood aswell as no touching anything outside the fumehood. Residues of hutners trace elements, which contained the heavy metals cobalt and copper(II) were disposed in a waste container for heavy metals. Common sense was also used.
Safe Shipment
Text goes here