Team:UANL Mty-Mexico/Safety/project

Would any of your project ideas raise safety issues in terms of researcher safety, public safety, or environmental safety?

To start, all persons within UANL_Mty iGEM team have gone through a course / required subject at the Universidad Autónoma de Nuevo León with duration of one semester, we have seen the protocols, rules, risk groups and how to respond to any contingency. During work lab with DNA’s and other BioBricks parts, we used only the organism E. coli K-12 sub-strains, which are considered risk level 1 according to the World Health Organization (WHO 2004).

For this year, Ecologic project is working with some new biobrick parts:

AIDA-I

Membrane adhesion protein obtained synthetically but originated from enteropathogenic E. coli, by itself no toxicity or pathogenicity is produced.

MOCR

Is a fusion tag protein mutant from Ocr bacteriophage T7 protein by substitution of two amino acids (DelProposto 2009) , used to increase the solubility of the protein associated and obtained synthetically.

RHMT

Is an arsenic human chelator protein, which has no toxicity or pathogenicity.

Besides using existing BioBricks:

K190028

K325101

J33201

Obtained from the registry of biological parts as non-hazardous.

Finally, consumption of water with arsenic doses greater than 0,025 mg / L are prohibited in Mexico by the NOM-127-SSA1-1994-2000 (SSA 2000) , due by human health damage.

It has seriously considered whether to use this year project, arsenic biosensor, chelator and remover outside the lab, so far we have decided just perform measurements and laboratory trials for this iGEM competition, we are working on placing biological locks to prevent damage to the environment if it gets released either accidentally or intentionally: e.g. an apoptosis mechanism (electric or magnetic) working in synergy with the high rate bacterial recovery system, using a membrane protein silica binding (AIDA). Additionally to confine bacteria that previously recovered arsenic, failing to return to the environment, and giving a future treatment in order to reinstate in a process of industrial interest.

For last year project which is going on HuBac, the parts BBa_K566000, BBa_K566001, BBa_K566002 were sent to synthesize and the sequence was obtained from phage lambda genome; these genes and composite parts were only used for regulatory purposes, while, not using the phage directly in the laboratory, the obtained genes are not pathogenic or toxic. We also worked with the Mnt repressor, BBa_K566008 from bacteriophage P22 that likewise does not present toxicity or pathogenicity. We also worked with PenI repressor synthesized from Bacillus licheniformis, which is considered risk level 1 organism (NIH) . Worked with the parts BBa_K566021, BBa_K566022, BBa_K566025, BBa_K566027, BBa_K566028 and derivatives from Synechocystis sp, that in most of the literature is considered as non-toxic, these worked genes were provided gently by Dr. Jeff Tabor from Rice University and previously tested experimentally in E. coli. Also worked with the fluorescent proteins GFP, RFP and CFP, which have already been tested in E. coli previously and are not considered as a risk.

For the use of chemical reagents and laboratory equipment we followed the biosecurity rules imposed by Microbiological and Biomedical Laboratories(HHS 2009) and the WHO (WHO 2004) , conducting standard microbiological techniques and practices, always using safety equipment and personal protective materials as lab coat, gloves, closed shoes, long pants, washing hands regularly, centrifuge safety lid, UV eye protection, restricted access to the lab area, reserved areas and adequate signs, etc…

If our project accidentally release into the environment, it would not respresent any problems with safety and public health, as all genes are natural to E. coli, that is world widespread and it is not pathogenic, besides being genetically weakened to die if not found in an environment with all the essential nutrients provided in a laboratory. For this, before disposing the laboratory resources, materials are pre-sterilized and only the people involved in the project can work with the biological parts and laboratory equipment, iGEM members have already been previously trained to avoid these situations happen. To prevent malicious use of the BioBricks parts and organism used, it has been seriously spoken with all the team members about the benefits / advantages as well as the dangers of this type of activities, so that everyone is aware of the magnitude of the events worked and agreed not to make improper use of the biological material.

Summarizing neither BioBricks used threatens team members, general public or the environment. Although protocols and biosafety rules are followed to avoid any possible contingencies.