Team:Freiburg Bioware/Safety
From 2010.igem.org
Biosafety
Legal regularisation in the Federal Republic of Germany
In Germany all working that includes recombinant DNA technologies is regulated by the Gesetz zur Regelung der Gentechnik. This law regulates general aspects arising from the life sciences and refers for more precise interpretations in §4 to the Zentrale Kommission für die Biologische Sicherheit. The ZKBS is a commission composed of 20 technical experts that releases yearly statements to actual issues of biosafety. So far the ZKBS released three stratements affecting the work with Adeno-associated viral systems 26, 27, 28. These documents were used to assess the dangers that could arise from our project to team members and the enviroment.
At the Albert-Ludwigs-University Freiburg for all concerns of security the Stabsstelle Sicherheit is responsible and to contact if questions arise. Especially for questions of biological security Dr. Petra Markmeyer-Pieles is cognizant. We contacted her a first time befor the begin of our project in March when it was clear that the Adeno-associated Virus (AAV-2) was chosen as the topic of our project. At that time she proposed to do the cloning in the AAV-2 that is for sure to handle under biological security level 1 and to prepare everything for work under biological security level 2 to satisfy the precaution principle. The precaution principle was realized and all viral vectors that contained a modified capsid were handled under SII conditions until proven harmless. In August the planing of the project was completed, summarized in an Biosafety application30 and handed to the department for biological security who approve the application in an official BSL1 confirmation31official BSL1 confirmation for our project.
Our project was designed in a way that it avoids any serious safety issues as far as possible. When working with infectious particles a minimal risk for the researcher is allways present. This risk was minimized by restricting the transduced genes to fluorescent proteins and prodrug convertases that are already proven not to harm human cells in the absece of the corresponding prodrug. A potential danger for the public or the environment was minimized as much as possible by following strictly the rules of Good Laboratory Practice (GLP) and the abdication of using randomized insertions in the capsid and of replication potent viruses. Minimizing the risk for team members and the society was was allways one of the major concerns, especially because worries about undergraduate students manipulating a virus could arise. The security concept will be explained by quoting and explaining the six guiding principles for safe manipulation of Gene Manipulated Organisms (GMOs) as summarized in Kimman et al. ; 200818.
Several composite parts that were assembled by our Team this year are alone capable of producing infectious viral particles when transduced together with a vector plasmid and a helper plasmid into AAV-293 cells. These special cells provide the adenoviral gene E1 stabily integrated in trans. These cells are not provided in the Virus Construction Kit nor availible in the Parts Registry and have to purchased from other laboratories or a commercial supplyer. For this reason we estimate the risk of a accidental transformation of AAV-293 cells with all three plasmids for negligible. Nevertheless we considered it useful to mark every BioBrick or Composite Part in the Registry that contributes to the production or is capable of producing viral vectors when transformed under the previously mentioned conditions.
psychological research into the concept of "identity-driven decision-making" (Torpman,2004) 19
Every grout has a set of norms: a code of conduct about what is acceptable beahviour (Jaques, 2004] 19
Trade-off between potential misuse and promising medical progress
In principle each research-project that bears any risks for engaged researchers, mankind or the environment should be treated under the precautionary principle as proposed 11: "treat synthetic microorganisms as dangerous until proven harmless".
This would mean to work on such synthetic DNA containing Bio Bricks at least under Biological security levels two.
Additional to this secure working environment the system itself can be optimized according to biosafety aspects, means to reduce it's viability outside the laboratory. This aim can be approached by reducing the systems ability to evolve, proliferate and interact with it's environment. A common method to achieve this goal is to engineer microorganisms in a way that they depend on nutrients that can't be found in the environment in sufficient amount.
Biosecurity
Def: "measures focus on the prevention of theft, misuse , or intentional relese of pathogens and toxins" 1Include something in your project description and presentations that demonstrates that you have thought about how others could misuse your work.For sure there is allway the possibility that knowledge to produce transgene viral vectors could be used to produce bioweapons. Therefor it was important for us to use a system that does not bear the risk that someone could use it for evil purpose. In the case of the Adeno-associated virus the very limited packaging capacity is the major reason that excludes it from the list of agents that could realistically be used for the pruduction of bioweapons. Even a fully replication potent AAV will depend on the coninfection of a helpervirus and is therefore not suitable for a fast propagation in an population. Additional to this point we concentrated our project on the retargeting of the virus - means to make the broad tropismn more narrow and to decrease the transduction efficiency in the most cases. This modification is usually mainly required for medical purposes. Also we did neither investigate possibilities to shield the vector from the immune system of potential host nor ways to bypass an existing immunity.
01 Synthetic Biology & Biosecurity - Awareness in Europe | Kelle ; 20071 02 Synthetic Biology - Social and Ethical Challenges | Balmer et Martin ; 20082 03) Synthetic Biology - Applying Engineering to Biology
used04 The Impact of the Development of Modern Biology and Medicine on the Evolution of Offensive Biological Warfare Programs in the Twentieth Century | Dando ; 1999 04 05 Chemical Synthesis of Poliovirus cDNA: Generation of Infectious Virus in the Absence of Natural Template | Cello et al. ; 2002 05 06 Dangerous research : When science breeds nightmares | Steinbruner et Harris ; 2003 06 07 Synthetic Genomics - Options for governance | Garfinkel et al. ; 2007 07 08 Synthetic biology | Benner et Sismour , 2005 08 09 Synthetic biology—putting engineering into biology | Heinemann et Panke ; 2006 09 10 Revealed: the lax laws that could allow assembly of deadly virus DNA | Randerson ; 2006 10 11 The Promise and Perils of Synthetic Biology | Tucker & Zilinskas ; 2006 11 12 1918 Flu and Responsible Science | Sharp ; 2005 12 13 The Darker Bioweapons Future | CIA ; 2003 13 14) [[Media:Primer for Synthetic Biology - Mohr 2007.pdf]] 15) [[Media:Freiburg10_The bugs of war.pdf]] 16 Expression of mouse interleukin-4 by a recombinant ectromelia virus suppresses cytolytic lymphocyte responses and overcomes genetic resistance to mousepox. | Jackson et al. ; 2001 16 17 Governance of dual-use research: an ethical dilemma. | Selgelid ; 2009 17 18 Evidence-based biosafety: a review of the principles and effectiveness of microbiological containment measures. | Kimman et al. 2008 18 19) A Hippocratic Oath for life scientists | Revill et Dando ; 2006 19 20 Empowerment and restraint in scientific communication. New developments make it easier to share information, but more difficult to deal with dual-use biology. | Campbell ; 200620 21 When risk outweighs benefit | Aken ; 2006 21 22 Advances in life sciences and bioterrorism. Risks, perspectives and responsibilities.| Beck ; 2003 22 23 PNAS policy on publication of sensitive material in the life sciences | Cozzarelli ; 2003 23 24 Characterization of the Reconstructed 1918 Spanish Influenza Pandemic Virus | Tumpey et al. ; 200524 25 Deadly flu virus can be sent through the mail| v. Bubnoff; 200525 26 Risk assessment of human Adeno-associated viruses| ZKBS; 200126 27 Advises for AAV carrying cell cycle regulating genes| ZKBS; 2004 27 28 Risk assessment of human Adeno-associated viruses and AAV derived vectors| ZKBS; 2005 28 29 Evidence for gene transfer and expression of factor IX in haemophilia B patients treated with an AAV vector.| Kai et al. ; 2000 29 Bioverteilung in Klinischer Studie 30 Biosafety application of the iGEM team Freiburg_Bioware 201030 (in German) 31 Official classification as Biological Safety Level 1 by the local biosafety office31