Team:UANL Mty-Mexico/Safety

Safety

We know biosafety measures must be inherent to any synthetic biology project. Through this questions we hope to solve any doubts concerning our project's safety:

1. Would any of your project ideas raise safety issues in terms of: researcher safety, public safety, or environmental safety?
2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?
3. Is there a local biosafety group, committee, or review board at your institution?
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?

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 substrains, which are considered Tier 1 risk group according to the WHO.

For this year, Ecologic project is working with some new biobrick parts: AIDA-I an 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 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 and J33201 obtained and presented in the registry of estándar biological parts as non-hazardous. Finally, the arsenic doses worked greater than 0,025 mg / L are prohibited in Mexico by the NOM-127-SSA1-1994-2000 causing human health damage.

It has seriously considered whether to use this year project, arsenic biosensor, chelator and remover in the environment, so far we have decided just perform measurements and laboratory trials at this iGEM competition, we are working on placing biological locks preventing 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 from phage lambda, 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 level 1 organism. 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 are considered as non-toxic, these worked genes were provided gently by Dr. Tabor 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

 and the World Health Organization, conducting standard microbiological techniques and practices, always using safety equipment and personal protective materials as lab coat, gloves, closed shoes, pants, washing hands regularly, centrifuge safety lid, UV protection, restricted access door, reserved areas and adequate signs, etc…

If our project accidentally is released into the environment, it would not pose any problems with safety and public health, as all genes are within 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 anyway followed to avoid any possible contingencies.

Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? If yes,

did you document these issues in the Registry?

how did you manage to handle the safety issue?

How could other teams learn from your experience?

The project of this year as well as the past, are based mainly on the use nontoxic/pathogenic parts. Although working with rare BioBrick derivate parts such as Mocr from Ocr (potent inhibitor of DNA restriction and modification enzymes type I of bacteriophage T7), no longer presents these effects because it have been modified in two amino acids. Also many parts used from bacteriophages are mostly regulatory used and show no damage to team members, general public or the environment.  We worked with E.coli organism level 1, which are well-characterized and present minimal hazard to laboratory personnel or environment.

 

As mentioned above, is not intended to release the microorganism to the environment (biosensor, chelator and arsenic remover), because much more testing, evidence, techniques and equipment is required, only laboratory work will be done for this year. Although it is planned that in a future after having spent enough money and evidence to prove lack of pathogenicity or horizontal transfer of genes and biological locks functioning properly, there is the possibility of using bioremediation in water contaminated with arsenic in major cities of Mexico.

We commit to document and record all parts of this project with all necessary information and clearly specify the parts that may have a conflict with biosafety issues.

Is there a local biosafety group, committee, or review board at your institution?

If yes, what does your local biosafety group think about your project?

If no, which specific biosafety rules or guidelines do you have to consider in your country?

We are ruled by the following regulations according to the biological context in our country:

Ley de Bioseguridad de Organismos Genéticamente Modificados (Biosafety Law for Genetically Modified Organisms.) 

http://www.diputados.gob.mx/LeyesBiblio/pdf/LBOGM.pdf

Reglamento de la Ley General para la Prevención y Gestión Integral de los Residuos. (General Law for Residues Prevention and Comprehensive Management's Guideline.) http://www.diputados.gob.mx/LeyesBiblio/regley/Reg_LGPGIR.pdf

Ley General del Equilibrio Ecológico y la Protección al Ambiente (General Law for Ecological Balance and Environmental Protection.) 

http://www.diputados.gob.mx/LeyesBiblio/pdf/148.pdf

Guía sobre la reglamentación relativa al Trasporte de sustancias infecciosas. (Guidance on regulations for the transport of infectious substances.)

http://www.who.int/csr/resources/publications/biosafety/WHO_HSE_EPR_2008_10_ES.pdf

Guía de evaluación de riesgos microbianos. (Guide microbial risk assessment.)

http://www.epa.gov/raf/files/mra-guideline-july-final.pdf

Manejo de Residuos Peligrosos Biológico-Infecciosos. (Biological and Infectious Hazardous Waste Management.)

http://www.uv.mx/vincula/vin_social/solicitudes/documents/ManejoResiduosPeligrosos.pdf

Manual sobre bioseguridad en el laboratorio, Organización mundial de la Salud. (Handbook of laboratory biosafety, World Health Organization.)

http://www.who.int/csr/resources/publications/biosafety/Biosafety7.pdf

NORMA Oficial Mexicana NOM-087-ECOL-SSA1-2002, Protección ambiental - Salud ambiental - Residuos peligrosos biológico-infecciosos - Clasificación y especificaciones de manejo. (Mexican Official Standard NOM-087-ECOL-SSA1-2002, Environmental Protection - Environmental Health - Hazardous biological and infectious waste - Classification and management specifications.)

http://www.salud.gob.mx/unidades/cdi/nom/087ecolssa.html

NORMA Oficial Mexicana NOM-017-STPS-2008, Equipo de protección personal-Selección, uso y manejo en los centros de trabajo. (Mexican Official Standard NOM-017-STPS-2008, Personal protection equipment - selection, use and management in the workplace.)

http://salud.chiapas.gob.mx/doc/biblioteca_virtual/normas/NOM-017-STPS-2008.pdf

Our project is currently within the analysis of the biosafety committee at the Universidad Autónoma de Nuevo León headed by Dra. Lydia G. Rivera Morales certified by the Mexican Association of Biosafety and Biosecurity (amexbio.org), and therefore we promise to post detailed comments later when we obtain them. Dra. Lydia, mentioned in advance to keep following biosafety precautions levels 1, 2 and materials properly disposals, finally commented in the recovery module by silica binding the consideration of biological locks in case of accidental releases. 

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?

Yes, in fact we can think of several ideas that could be consider for future iGEM competitions, one of them is an essay required for all participating teams about their project concerning synthetic biology, biosafety and ethics, regardless of whether or not the project has major biological problems. It will require not only thinking and reflection on all the issues involved, and will help them answer the safety questions more accurately and appropriately; this essay would aware iGEM students what they are creating and that entails a synthetic biology project.

We propose an implementation or creation of new software, where each iGEM teams should run the software analysis and detection tools for toxicitivity or pathogenic criteria in all their BioBricks, and clearly post the test result to the wiki’s section of biosecurity. This way, if a team is performing a dangerous part, the analysis result would show a warning sign and they could be informed in advance, placing the record in a visible part of the wiki and the relevant information related to that part in the registry part, otherwise disposing of part or discontinue working on it

Last but not least, another way but slightly futuristic is start using XNA "xeno-nucleic acid", iGEM teams could use different molecules with similar functions that allow us store information, therefore there could be less risk of horizontal transfer or interference with other living organisms, while working with synthetic biology projects.