Ehime-Japan iGEM Team: Welcome


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

No. Our project ideas do not raise safety issues in terms of them. We used the light sensor genes from Synechocystis and the lon and the ssrAtag genes from Mesoplasma florum in and Also, the lux and las gene regularion systems were used in E.cold-heat sensing system. An explanation of the safety standard about our experiments is as follows.
In Japan, we must document our local committee which organism the gene is from and what the host isbefore we carry out gene recombination experiments. The organisms that the gene used in our projects are originated from are shown in Table1 with the Class as defined in Japanese ordinance that the organism are classified into.

Table1. The safety classes
The organism providing the genes Gene Class
Synechocystis sp.PCC6803 ccaS,ccaR,cph8,cpcG2promoter,ho1,andpcyA Class1
Mesoplasma florum L1 Lon protease,ssrAtag Class1
Vibrio fischeri LuxI,Luxr,and PLuxR Class1
Pseudomonas aeruginosa lasI,lasR,and PlasR Class2
Aequeora Victoria GFP Class1
Discosoma striata RFP Class1
Enterococcus faecalis Aad9 Class1

The hosts used in our experiments are shown in Table 2.
Table2. The safety classes of host
Host Originalstrain Class
E. coli DH5α K12 Class1
E. coli JT2(ΔEnvZ) K12 Class1
E. coli JW0429(Δlon) K12 Class1

※Class1: All the organisms expect class 2to4.
Class2:The organisms having low virulence to mammals.
Classes3and4 can cause more risks.
The organisms we are working with are E.coli strains DH5α,JT2(ΔEnvZ),and JW0429(Δlon)and they have no virulence. Thus,they are all in Class1.According to the Japanese regulations, the containment measure(measure to be taken to prevent disposal of LMOs)to be taken should be determined primarily considering the Classes of the donor and host organisms.However,if the gene to be inserted into the host is already characterized to have no relation to the virulence or transmissibility of the organism from which the gene is originated,then,the containment measure can be determined only from the host Class.

In our case,Pseudomonas is in Class2,but the las regulation system by itself has no relation to the virulence:the system does not contain the lasBgene,which is regulated by the las system in the organism and causes the virulence.We also planned to use the samples from Mycoplasma capricolum,which are in Class2,though we did not use them. The situations are the same as in the case of Pseudomonas. All the other genes also have nothing to do with virulence to mammals. Therefore,the level of the containment measure to be taked to prevent dispersal of LMOs is P1,as determined from the Class of the host.Our documents were examined in our local committee and accepted without rasing any problem.Thus,we are doiing all experiments under the condition of P1 level experiment.
Table3.The P1 level containment measures to prevent dispersal of LMOs.Wecan conclude,therefore,that our project ideas are safe for researcher,public,and the environment.

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?

No. Our new BioBrick parts are safe and all the genes we used are BL1 or BL2.

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? If no, which specific biosafety rules or guidelines do you have to consider in your country?

Yes. Before we started the experiments, we submitted the safety list about our projects to our biosafety committee, and it was accepted without any problem pointed outs.

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?

We think it would be good to make a check list in order to confirm whether projects or experimental equipment are proper. Also, in case of using a virulent organism or a gene from it, the user should show clearly how to deal with it. We can easily answer and find some problems we did not think.