Team:Freiburg/Safety

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Safety

General biosafety regulation in Germany

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

In Germany all work that includes recombinant DNA technologies is regulated by the "Gesetz zur Regelung der Gentechnik". This law regulates general aspects in 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.

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 Max Planck Institute for Immunobiology and Epigenetics, Freiburg, for all concerns of security the Arbeitssicherheit is responsible and to contact if questions arise. Especially for questions of biological security Dr. Siegfried Kühn is cognizant. We contacted him the first time before the start of our project in March when it was clear that we were going to use a lab in the Max Planck facility. We got the permission for S1 work, but with a restriction to use only E. coli and not e.g. yeast, since the institute does not want to have spore making organisms in the building, to prevent uncontrallable contaminations. All our used equipment, chemicals, genetic parts and organisms were first approved by the Arbeitssicherheit.

Risk management

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. We do not use any human related sequences or that of any pathogens. Our used E. coli strains are BL21 and a derivate of E. coli K12.

1) Hazard recognition and identification Risk assessment has been done and all legal regulations were considered.

2) Biological containment: Biological containment means the usage of organisms with "reduced replicative capacity, inefectivity , transmissibility, and virulence"18.

3) Concentration and enclosure: all work was done in a S1 lab inside the Max Planck institute certified by the German government.

4) Exposure minimization: Labcoats were worn, food and drinks were only allowed outside the lab. We furthermore eliminated all toxic reagents in the lab, except acids and bases, which we stored separately in special shelfes to contain them. We replaced Ethidiumbromide by a non toxic surrogate, which is proven to be harmless in mammals.

5) Physical containment: The lab is physically separated by glass doors to the hallway. The doors were always locked when nobody was in the room.

6) Hazard minimization: We avoided to use hazardous protein sequences on purpose.

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?

We carefully avoided to use any pathogenic or hazardous sequences in our de novo protein design. However this needs special attention, since unwatchfully copying adequate sequences from databases sometimes leads to at least partial pathogenic protein designs. In some databases these dangerous sequences are not outlined as such and it needs further investigation to find out about their natural functions. We did this and therefore managed to avoid copying a sequence of a protein from yersinia pestice.

1. Would the materials used in your project and/or your final product pose:

a. Risks to the safety and health of team members or others in the lab?

No, because we are using safe E. coli strains that are not viable outside the lab. The host strains are not pathogenic, infectious or toxic. We do not intent to express proteins that increase the strains pathogenicity, infectivity or toxicity. We furthermore eliminated all toxic reagents in the lab, except acids and bases, which we stored separately in special shelfes to contain them.

b. Risks to the safety and health of the general public if released by design or accident?

No, the strains used are non-infectious or pathogenic. Since the strains are not viable outside the lab environment and we do not intend to increase their abilities to survive no risk to public health and safety.

c. Risks to environmental quality if released by design or accident?

No, since the strain is not viable outside the lab and we do not intent to express proteins that increase its viability, it poses no threat to the environment.

d. Risks to security through malicious misuse by individuals, groups or states?

As any protein expression vehicle, our product can be used to express malicious proteins, like proteinous toxins or proteins of a synthetic pathway to produce other toxic substances. To prevent misuse by individuals, groups or states we plan to ship the product only to known institutes or recipients. To prevent security issues, the lab and storage rooms are allways locked when left, to assure that non of our work can by taken and used witout oure knowledge.

2. If your response to any of the questions above is yes:

a. Explain how you addressed these issues in project design and while conducting laboratory work.

b. Describe and document safety, security, health and/or environmental issues as you submit your parts to the Registry.

3. Under what biosafety provisions will / do you operate?

a. Does your institution have its own biosafety rules and if so what are they? Provide a link to them online if possible.

The institutes of the Max Planck Gesellschaft operate with the national German biosafety regulations that can be found e.g. under the following link (german): http://www.rp.baden-wuerttemberg.de/servlet/PB/show/1103199/rpk24_laborrichtlinien.pdf

b. Does your institution have an Institutional Biosafety Committee or equivalent group? If yes, have you discussed your project with them? Describe any concerns or changes that were made based on this review.

At the Max Planck Institute for Immunobiology and Epigenetics, Freiburg, for all concerns of security the Arbeitssicherheit is responsible and to contact if questions arise. Especially for questions of biological security Dr. Siegfried Kühn is cognizant. We contacted him a first time before the beginning of our project in March when it was clear that we were going to use a lab in the Max Planck facility. We got the permission for S1 work, but with a restriction to use only E. coli and not e.g. yeast, since the institute does not want to have spore making organisms in the building, to prevent uncontrallable conaminations. All our used equipment, chemicals, genetic parts and organisms were first approved by the Arbeitssicherheit. The projects have been discussed with the person in charge and no changes have been introduced to the project or materials used in the project.

c. Will / did you receive any biosafety and/or lab training before beginning your project? If so, describe this training.

The members of the team all have taken courses in different laboratories, and in each course biosafety instructions are part of the lab trainings. In addition, we had a short introduction in general lab and biosafety by the institutes’ officer of safety.

d. Does your country have national biosafety regulations or guidelines? If so, provide a link to them online if possible.

In Germany all work that includes recombinant DNA technologies is regulated by the Gesetz zur Regelung der Gentechnik. This law regulates general aspects in 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.

http://www.gesetze-im-internet.de/gentg/
http://www.gesetze-im-internet.de/bundesrecht/gentg/gesamt.pdf

@@ Line 27: / Line 27: @@
 :::::Our project was designed in a way that it avoids any serious safety issues. We do not use any human related sequences or that of any pathogens. Our used ''E. coli'' strains are BL21 and a derivate of ''E. coli'' K12.
 :::::1) Hazard recognition and identification Risk assessment has been done and all legal regulations were considered.
-:::::2) Biological containment Biological containment means the usage of organisms with "reduced replicative capacity, inefectivity , transmissibility, and virulence"18.
+:::::2) Biological containment: Biological containment means the usage of organisms with "reduced replicative capacity, inefectivity , transmissibility, and virulence"18.
-:::::3) Concentration and enclosure All work was done in a S1 lab inside the Max Planck institute certified by the german government.
+:::::3) Concentration and enclosure: all work was done in a S1 lab inside the Max Planck institute certified by the German government.
-:::::4) Exposure minimization Labcoats were worn, food and drinks were only allowed outside the lab. We furthermore eliminated all toxic reagents in the lab, except acids and bases, which we stored separately in special shelfes to contain them. We replaced Ethidiumbromide by a non toxic surrogate, which is proven to be harmless in mammals.
+:::::4) Exposure minimization: Labcoats were worn, food and drinks were only allowed outside the lab. We furthermore eliminated all toxic reagents in the lab, except acids and bases, which we stored separately in special shelfes to contain them. We replaced Ethidiumbromide by a non toxic surrogate, which is proven to be harmless in mammals.
-:::::5) Physical containment The lab is physically separated by glass doors to the hallway. The doors were always locked when nobody was in the room.
+:::::5) Physical containment: The lab is physically separated by glass doors to the hallway. The doors were always locked when nobody was in the room.
-:::::6) Hazard minimization We avoided to use hazardous protein sequences on purpose.
+:::::6) Hazard minimization: We avoided to use hazardous protein sequences on purpose.
@@ Line 38: / Line 38: @@
 '''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?'''
-:::::We carefully avoided to use any pathogenic or hazardous sequences in our de novo protein design. However this need special attention, since unwatchfully copying adequate sequences from databases sometimes leads to at least partial pathogenic protein designs. In some databases these dangerous sequences are not outlined as such and it needs further investigation to find out about their natural functions. We did this and so managed to avoid copying a sequence of a protein from yersinia pestice.
+:::::We carefully avoided to use any pathogenic or hazardous sequences in our de novo protein design. However this needs special attention, since unwatchfully copying adequate sequences from databases sometimes leads to at least partial pathogenic protein designs. In some databases these dangerous sequences are not outlined as such and it needs further investigation to find out about their natural functions. We did this and therefore managed to avoid copying a sequence of a protein from yersinia pestice.