Team:GeorgiaTech/Safety

Safety

Safety is an important factor while conducting a lab experiment involving synthetic biology and utmost care should be taken such that any accidental or experimental mistakes do not harm the welfare of staff as well as the community. That is why, at Georgia Institute of Technology, safety procedures are in place to minimise the risk of mishaps in a laboratory.

General Lab Safety: The Georgia Tech iGEM 2011 team follows the guidelines as implemented by the university and all of its members have been trained in Lab Safety Protocols for this purpose. All the team members have been trained by the lab safety officer in School of Biology, Mark Pline by conducting a presentation on general safety in the lab, understanding material safety data sheets, the meaning of various warning labels, such as those on refrigerators, waste disposal, and autoclaving techniques. The GT iGEM team follows all the safety protocols and guidelines as described by the Georgia Tech Environmental Health & Safety here: http://www.ehs.gatech.edu/

Also, our project follows the policies and procedures governing the possession and use of Recombinant DNA as laid out by the Georgia Institute of Technology Institutional Biosafety Committee and as laid out in the document attached here: http://www.compliance.gatech.edu/forms/IBC/IBC_PP.pdf

Safety Issues Concerning the GT iGEM 2011 CRISPR Project:

Environmental and Public Safety:

Bacterial Strains Used:

1) Streptococcus thermophilus DGCC 7710: Our project involves the use of Streptococcus thermophilus, a gram-positive bacteria that is not known to cause disease in humans. Although other members of its genus are pathogenic, S. thermophilus is completely safe, growing naturally in the human intestines, and is in fact added to yogurt as a probiotic as indicated in the following link : (http://www.nature.com/index.html?file=/nbt/journal/v22/n12/full/nbt1034.html). 2) Bacillus subtilis 168: Another bacteria we are using, Bacillus subtilis, is classified as a Safety Group 1 organism by the National Institutes of Health, thus an “agent that is not associated with disease in healthy adult humans.” It has also been evaluated by the Environmental Protection Agency, and their conclusions were that it is not considered pathogenic to humans, animals, or plants as indicated here: (http://epa.gov/biotech_rule/pubs/fra/fra009.htm) 3) Escherichia coli BL21: The bacteria we used, BL21 electrocompetent E. coli, are non-pathogenic and unlikely to survive outside of a laboratory setting, thus not causing any ill effects if released by accident.

Safety Concerns Concerning the CRISPR mechanism:

The Georgia Tech iGEM project involves utilizing CRISPR systems to decrease antibiotic resistance in bacteria, with possible future applications in human health. The CRISPR system is naturally prevalent in 40% of the bacterial population and hence it is highly unlikely that the genes under experimentation would cause adverse effects beyond what is already present in nature. The CRISPR system acts by increasing the resistance of the bacteria against plasmid uptake and phage attacks, which is unlikely to cause increased pathogenicity of the bacteria. Rather, since plasmids in nature can harbor antibiotic resistance genes or other genes that enable a bacterium to survive and propagate, decreasing plasmid uptake may actually decrease pathogenticity. Similarly, CRISPR’s resistance mechanism works only against nucleic acids, so adding it to new species will not enable that species to acquire resistance to antibiotics - which are typically small molecules, not nucleic acids. For the purposes of general safety, the manipulated CRISPR system will only target genes that have been specifically engineered for targeting and precautions have been taken to ensure that none of these should be detrimental to the community if released by accident.

Security Risk ?:

Our project promises of developing a CRISPR mechanism which could eliminate resistance from the inserted bacteria against a certain already present resistance. Although there is a potential to misuse any genetic modification experiment, the experimental product from this project will be highly regulated and will not pose any new security threat.

Biobrick Safety Considerations:

Our submitted biobrick device will consist of the CRISPR locus taken from S. thermophilus. As the submitted biobrick will have a manipulated CRISPR mechanism which will only target the spacers inserted by our team, it will not pose any safety risk as it will only carry out its intended purpose.

Biosafety Regulations at Georgia Institute of Technology:

Our project has been classified as biosafety level 1 as all of our organisms are “unlikely to cause human diseases” and the Occupational Health and Safety Committee and the Environmental Health and Safety Office of the Georgia Institute of Technology has approved Professor Eric Gaucher’s laboratory (the lab under which the GT iGEM 2011 team works) for Biosafety Level 1 Research and no concerns have been raised by the committee as regards to our project. As stated above, we have been in constant touch as regards to general lab safety with our department’s lab safety officer Mark Pline and have adhered to all th basic safety protocols. Also, our project follows the guidelines laid out by the national safety guidelines as laid out by CDC here http://www.cdc.gov/biosafety/publications/bmbl5/index.htm.

Also, as stated above in General lab safety, our project follows the policies and procedures governing the possession and use of Recombinant DNA as laid out by the Georgia Institute of Technology Institutional Biosafety Committee and as laid out in the document attached here: http://www.compliance.gatech.edu/forms/IBC/IBC_PP.pdf

As a final checkpoint, our proposal will be consistently and constantly verified in detail to ensure that every aspect complies with university and state safety regulations

Combined efforts of all iGEM teams to ensure safety: Our group has discussed the sharp learning curve associated with having a young iGEM team with relatively little lab experience among them. All group members attended an institute biological laboratory safety class, but many things were learned along the way, specific to genetic engineering research. We would propose a collaboration of iGEM members to produce an online, visual, one-hour introductory course on the basic biosafety issues that could arise during the iGEM research process.