SAFETY AND SECURITY QUESTIONS

GENERAL

The strains we utilized are Escherichia coli. DH5α, BL21 and ΔCheZ, a cheZ gene knocked out strain modified from BL21. These microbes which have been proved pathogenicity-free are very common in labs and have been used in our lab for many years, and therefore the protocols for dealing with the biological waste are comprehensive. (the highlights of the protocols will be shown in the followings and the complete bio-safety operation manual is ready for downloading in PDF format)

All the genes (recAp, recNp, cheZ, and trkD) we manipulated are constructively internal genes of E. coli, we have not transferred any external gene into any microbes we have used. So the potentially poisonous products are unlikely to be produced.

All the reagents we have used are common reagents for molecular biological experiment, and have been properly disposed by a professional bio-waster-dealing company.

All the work already done yet is restricted in laboratory. However, if our bacterial nuclear leakage rescuer is to be used in field, we will use a device rather than just pour the bacteria into open water to minimize the hazard of bio-safety issues. The possibility that the bacteria will be eaten by fish is factored into our design of the device. You can see our AGGREGATION & RECOVERY part under the PROJECT section to find out more details about our devices.

Most of our experiments were conducted in Microbial Genetics Laboratory (MGL) advised by Prof. Yongjun Lu. MGL is a Bio-safety Level 1 (BSL-1) laboratory and is suitable for experiments dealing with well characterized reagents including non-pathogenic Escherichia coli. not known to cause diseases. Standard protocols for operation with microbe will provide basic protection to operators.

SAFETY EQUIPMENT

Two superclean benches are equipped for normal microbiological operation.

Doors with Access-Control-By-ID-card System are equipped to prevent un-authorized entering both for human and the other animals that are potentially possible to carry out reagents from laboratory.

Windows with screens to prevent insects and birds from entering the laboratory

Rubber gloves for hand protection and glassware for eye protection.

STANDARD OPERATION

(defined by Biosafety Committee of Sun Yat-Sen University )

Everyone is restricted to the laboratory without the permission from the director of the laboratory. All children will not be allowed to enter the laboratory.

Bio-safety figure 1 (BSF-1) with laboratory director's name and contact is attached on the doors.

The doors should be closed.

No animals are allowed in the laboratory.

People conducting experiments must put on gloves and wear uniforms which should be stored separately.

Eating, drinking, smoking, handling contact lenses, applying cosmetics, storing food and drink is not permitted.

Mouth pipetting is prohibited, we use mechanical pipetting instead.

Used containers, samples, medium must be sterilized (e.g. autoclaving) before recycle and disposal.

Wastes must be decontaminated first before disposal, and different wastes must be disposed with different waste-disposal-systems according to risk evaluation.

Plans limiting arthropods and rodents are created and performed.

Everyone in the laboratory are well instructed the bio-safety operation manual and tested.

 

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?

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

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

d. Risks to security through malicious misuse by individuals, groups or states? Please explain your responses (whether yes or no) to these questions. Specifically, are any parts or devices in your project associated with (or known to cause):

- pathogenicity, infectivity, or toxicity?

- threats to environmental quality?

- security concerns?

NO.

a. The reagents we used were all common ones and had been proved non-pathogenic, non-infective and non-toxic. And Escherichia coli DH5α, BL21 and ΔCheZ strains we utilized are well-known non-pathogenic, non-infective and non-toxic ones too. Besides, the genes we manipulated were all internal, and thereby unlikely to raise environmental issues. By strictly abiding by the Bio-safety Operation Manual defined by Biosafety Committee Of Sun Yat-Sen University, we were well protected.

b. As mentioned above, all the materials were non-pathogenic, non-infective and non-toxic, and unlikely to cause diseases. And since we followed the rules defined by Bio-safety Operation Manual, it was unlikely that the materials would be released to publics.

c. The amount of genetically modified organism is very little in the scale of "environment" and it is environmental friendly as mentioned above. So, if released by design or accident, though the possibility is small, our genetically modified organism is not to risk environmental quality.

d. We have strictly abided by the Bio-safety Operation Manual, it is unlikely for our genetically modified organism to be released . Besides, the genetically modified bacteria are bio-safe. In all, with access-control system, it is difficult for malicious individuals to enter the laboratory and even though the bacteria do be released somehow, it will not risk the environment

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

a. Did you document these issues in the Registry?

b. How did you manage to handle the safety issue?

c. How could other teams learn from your experience?

No biobricks made by the SYSU-China team will raise any safety issues. The biobricks, such as recA, recN, cheZ, trkD and ag43, are all the natural parts of E.coli in natural environment, and commonly used for laboratory operation. Their function and functional mechanism are clearly known, and they are not associated with pathogenicity, infectivity, or toxicity, nor will they cause threats to environmental quality. Moreover, cheZ and ag43 are standard biobricks provided by iGEM authority, so the safety of these biobricks can be guaranteed.

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.

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.

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

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

a. SYSU_China team members abided strictly by the established safety rules in the lab (click here to download the rules in PDF format ) which was constituted by Biosafety Committee Of Sun Yat-Sen University on Dec. 12, 2007.

b & d. Second, In our university, Biosafety Committee of Sun Yat-sen University is responsible for monitoring the safety of all the researching activities on the campus. Their regulations are stipulated according to the WHO Laboratory biosafety manual, which is also conformed by our country (http://biosafety.sysu.edu.cn/Soft/UploadSoft/200804/2008042116071397.pdf). Before our team started the program, we had talked about the safety issues of the whole project. They were really concerned about the radiation-related experiments, and emphasized that these experiments must be operated by the professional technician. In addition to that, they also requested that all procedures conducted in this project should be performed according to the rules stipulated by WHO. On the whole, they considered our project as safe. With the surveillance of the Biosafety Committee of Sun Yat-sen University and the cautious operation of our members, the project will be harmless to the researchers, the public or the environment.

c. Each one in the lab was required to attend a pre-lab training led by advisors and graduate students both on experimental skills and safety instructions. The training included basic safety rules and skills in the lab, such as how to use the toxic reagents, how to deal with pathogenic organisms and etc. During the program, we wear gloves properly and stetilized tubes and plates and other containers after use.

4. OPTIONAL QUESTION: Do you have other ideas on how to deal with safety or security issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

We suggest that the safety information of any part used in the iGEM competition, especially its functions, should be investigated clearly through literature search before the usage. If the part has any intention to raise safety issues, the iGEM team should refuse to use it and choose other alternatives. In addition to that, iGEM teams should submit the safety report with their biobricks part to the iGEM authority, describing the safety issues of the biobricks part, which should also be examined by a professional third party. Thus the high-risk biobricks or parts with potential biosafety problems will be identified and abandoned before submitting. Through this way can the parts, devices and systems be made much safer. With above measures, the safety of the biobricks submitted can be guaranteed.