Team:Aberdeen Scotland/Safety

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SAFETY SOLUTIONS

Our iGEM team genetic engineering activities were regulated by our Institute's Advisory Committee on Genetic Manipulation (ACGM). We were required to submit a complete project description (see file at; media:Gmag_for_aberdeen.pdf) , which included a statement of the potential for the engineered organisms to cause harm, and precautions that would be taken to mitigate the risks involved with the project.

This document, and our discussions with the Advisory Committee on Genetic Manipulation required us to address the four safety questions posed as a condition of iGEM competition entry:
1. Would any of your project ideas raise safety issues in terms of researcher, public or environmental safety?
Our ACGM committee were satisfied, on the basis of our risk assessment document, that our project posed no such safety issues if the genetically manipulated organsisms created were restricted to lab use, and that all microbial cultures were killed before disposal (see below).

2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?
All of the parts in use by our team had previously been used as part of published molecular biology projects, and were considered to have no specific safety issues.

3. Is there a local biosafety group, committee, or review board at your institution?
Yes, the Advisory Committee of Gnetic Manipulation; our project was fully considered by this local biosafety group, on the basis of the assessment document we submitted (see linked pdf, above)

4. Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?
Our iGEM team this year and last year was required to provide interim experimental data to the ACGM committee to allow committee members to reach an assessment of possible emergent properties, and their associated risks, of the circuit we were constructing. In this way, any particular risks of a synthetic biology circuit operation could be mitigated using a process of ongoing and interim oversight, rather than the single safety assessment that an ordinary molecular biology project would be subject to.

Below, we outline some of the safety questions that our local ACGM committee required us to address;
From the genetic safety document, we were asked for instance to define; 'Potential for harm';
We stated; As commonly-used reporters, fluorescent proteins, translational repressors and native yeast enzymes, expression of the genes listed is unlikely in the extreme to endow either E.coli or S.cerevisiae with any harmful properties. .None of the gene products is known to be toxic. Antibiotic resistance markers, reporter genes, fluorescent proteins and RNA or DNA binding/repressor proteins listed likewise have no known harmful properties, and are in widespread established use in molecular biology.

From the genetic safety document, we were then asked; 'How might the GMM be a hazard to human health? Evaluate the severity of the harmful effects if they were to occur'
We stated; E. coli K12 and its derivatives are multiply disabled and are designated as Class 1 organisms. Good microbiological practice will be followed when using these organisms and over many years of use, no adverse effects have been noted.

Should transfer occur, the nature of all of the genes being manipulated (housekeeping or reporter genes) means deleterious consequences are unlikely in the extreme to result. Plasmids used are mobilisation defective. No genes are being expressed at unusually high levels, it it is considered very unlikely they will generate toxicity, in the extremely unlikely event of survival of the disabled E.coli host in the body, None of the genes that are to be propagated in E.coli are known to have deleterious effects on human health. All genes occur naturally in normal cells and are involved in basic metabolic processes. It is highly unlikely that even if large amounts of GMM were ingested that the protein would be targeted in sufficient quantity to a location likely to cause detrimental effects.

Saccharomyces cerevisiae strains almost exclusively carry multiple auxotrophic mutations and are designated as Class 1 organisms. Good microbiological practice will be followed when using these organisms and no adverse consequences have been reported from the genetic manipulation of this strain over many years of use in multiple laboratories.

There is extremely low likelihood of transfer of genes from S,cerevisiae to other micro-organisms. Should transfer occur, the nature of the genes being manipulated (housekeeping or reporter genes) means deleterious consequences are unlikely in the extreme to result. Plasmids used are mobilisation defective.

From the genetic safety document, we were finally asked; 'All GMMs in contaminated material and waste must be inactivated by "validated means", the method of inactivation chosen being appropriate to the level of risk.If chemical means of inactivation are to be used, what chemicals will be used and at what concentrations?'
We stated; 1 % Virkon (small volume spills). Virkon powder (large volume spills). 70% ethanol. Chemical means will only be used to disinfect surfaces and in the case of accidental spillage. Decontamination methods are specified in detail in our Local Rules for GM work.

From the genetic safety document, we were finally asked; 'What "degree of kill" is [autoclaving] expected to achieve? How has it been arrived at?'
We stated; 100%. Reasonable expectation of kill of microorganisms exposed to 126°C for 14 minutes (standard autoclave cycle, Prestige Autoclaves). While this autoclave cycle is non-standard (usual cycle is 121°C, 15 min.), our laboratory has experimentally tested the effectiveness of the Prestige cycle, and found it to completely kill E.coli cells, in standard 100 ml volumes in flasks placed in the middle of a full autoclave load.

In summary, the entire project was only initiated once a considered plan had been developed for safety management, and that plan approved by an independent committee of local experts who would give permission for practical work to begin once any safety concerns had been addressed

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