Team:Freiburg Bioware/Safety

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Biosafety

Legal regularisation in the Federal Republic of Germany

Which specific biosafety rules or guidelines do you have to consider in your country?

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.



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?

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.



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

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. Additionally all modified viral vectors were handled under biological safety level II. 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. Even if at none of the steps ever a serious safety risk was taken. The security concept will be explained by quoting and explaining the six guiding principles for safe manipulation of Gene Manipulated Organisms (GMOs).

  • 1) Hazard recognition and identification Risk assessment has been done and all legal regulations were considered as described in the last paragraph.
  • 2) Biological containment Biological containment means the usage of organisms with "reduced replicative capacity, inefectivity , transmissibility, and virulence"18. For our project only replicative deficient viruses were used, additional all modifications aimed to have a specific targeting of the resulting viral vectors for a specific cell type. This gain in specificity requires it to cut off the braod natural tropism resulting is an less infective virus copared to the wild type virus.
  • 3) Concentration and enclosure All working steps for the preparation of viral vectors were carried out in a BSL II laboratory and within this laboratory work with the AAV was restricted to a separate Laminar flow cabinet type II. Cell culture and storage of the virus was also done in separate Freezers and incubators. All laboratories and epipment that contained viral vectors were specially marked, for example with a Biohazard warning signs
  • 4) Exposure minimization This aspect of the guiding principles can be sumarized under "operator protection"18. The Exposure minimization was achieved in our laboratory by wearing special labcoats for the SII laboratory and gloves that were desinfected and changed regualrely. During manipulation of viral vectors attention was payed to avoid droplets and especially aerosoles. A possible diversion of the viral vectors was avoided by cleaning all equipment when inserted or removed from the Laminar flow cabinet and after completion of the work task.
  • 5) Physical containment The requirements for the physical containment were fullfilled by performing all manipulation on the AAV in an BSL II laboratory that guaranted a restriction of persons that entered the laboratory.
  • 6) Hazard minimization For the AAV-2 there are no sugestive activitis because the possible danger that runs out of the AAV is comparably low, vaccination is not avilible and biomonitoring is not necessary.
  • 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?

    Warning sign for part descriptions 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.

  • Description of the AAV-293 cell line
  • Contribute to community discussions on what needs to go into a code against the use of our science for hostile purposes (see A Community Response)

  • Public Perception
  • importance of maintaining public legitimacy and support
  • apllications should demonstrate clear social benefits
  • not overhyped - anxiety and unrealistic hopes
  • 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 - The Adeno-associated Virus as an example

    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?

    n principle each research-project that bears serious risks for mankind or the environment should be treated with the precautionary principle as proposed by 11: "treat synthetic microorganisms as dangerous until proven harmless" would mean to work on synthetic DNA containing Bio Bricks under Biological security levels three or four.
    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

    Include something in your project description and presentations that demonstrates that you have thought about how others could misuse your work.

  • 01) Synthetic Biology & Biosecurity - Awareness in Europe by Dr. Alexander Kelle in 2007 [http://www.idialog.eu/uploads/file/Synbiosafe-Biosecurity_awareness_in_Europe_Kelle.pdf Kelle ; 2007 1]
    used
  • 02) Synthetic Biology - Social and Ethical Challenges [http://www.bbsrc.ac.uk/nmsruntime/saveasdialog.aspx?lID=2277&sID=4233 2]
    used
  • 03) Synthetic Biology - Applying Engineering to Biology
    used
  • 04) The Impact of the Development of Modern Biology and Medicine on the Evolution of Offensive Biological Warfare Programs in the Twentieth Century [http://www.informaworld.com/smpp/content~content=a713604665&db=all]
  • 05) Chemical Synthesis of Poliovirus cDNA: Generation of Infectious Virus in the Absence of Natural Template [http://www.sciencemag.org/cgi/content/full/297/5583/1016 Cello et al. ; 2002 5 ] used
  • 06) [http://www.nytimes.com/2003/12/03/opinion/03iht-edstein_ed3_.html Dangerous research : When science breeds nightmares]
  • 07) [http://www.jcvi.org/cms/fileadmin/site/research/projects/synthetic-genomics-report/synthetic-genomics-report.pdf Synthetic Genomics - Options for governance]
  • 08) Synthetic biology by [Benner & Sismour 2005] [[Media:Freiburg10_Synthetic_Biology_by_Benner_and_Sismour.pdf | here]]
  • 09) Synthetic biology—putting engineering into biology by [Heinemann & Panke 2006]
  • 10) [http://www.guardian.co.uk/world/2006/jun/14/terrorism.topstories3 10] Revealed: the lax laws that could allow assembly of deadly virus DNA]
  • 11) [http://www.thenewatlantis.com/docLib/TNA12-TuckerZilinskas.pdf The Promise and Perils of Synthetic Biology [Tucker & Zilinskas, 2006]]
  • 12) [http://www.sciencemag.org/cgi/content/short/310/5745/17 1918 Flu and Responsible Science] Reaction to the H1N1 sequence [http://www.sciencemag.org/cgi/reprint/310/5745/17.pdf Sharp ; 2005 12] used
  • 13) [http://www.fas.org/irp/cia/product/bw1103.pdf The Darker Bioweapons Future by the CIA in 2003]
  • 14) [[Media:Primer for Synthetic Biology - Mohr 2007.pdf]]
  • 15) [[Media:Freiburg10_The bugs of war.pdf]]
  • 16) [[Media:Freiburg10_Expression of Mouse Interleukin-4 by a Recombinant Ectromelia Virus Suppresses.pdf]] [http://www.ncbi.nlm.nih.gov/pubmed/11152493 Jackson et al. ; 2001 16 ] used
  • 17) [http://www.ncbi.nlm.nih.gov/pubmed/19784453] [[media:Freiburg10_Governance_of_dual-use_research-_an_ethical_dilemma.pdf]]
  • 18) [http://www.ncbi.nlm.nih.gov/pubmed/18625678 18] [[Media:Freiburg10_Evidence-based_biosafety-_a_review_of_the_principles_and_effectiveness_of_microbiological_containment_measures.pdf]]
  • 19) [http://www.ncbi.nlm.nih.gov/pubmed/16819452 19] [[Media:Freiburg10_A Hippocratic Oath for life scientists.pdf]]
  • 20) [http://www.ncbi.nlm.nih.gov/pubmed/16819443] [[Media:Freiburg10_Empowerment and restraint in scientific communication.pdf]]
  • 21) [http://www.ncbi.nlm.nih.gov/pubmed/16819441] [[Media:Freiburg10_When risk outweighs benefit.pdf]]
  • 22) [http://www.ncbi.nlm.nih.gov/pubmed/12789408 22] [[Media:Freiburg10_Advances in life sciences and bioterrorism - Risks perspectives and responsibilities.pdf]]
  • 23) [http://www.ncbi.nlm.nih.gov/pubmed/12590130] [[Media:Freiburg10_PNAS policy on publication of sensitive material in the life sciences.pdf]]
  • 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