Safety
The Streptomyces chassis
Our project CO2CURE consists in developing a bacterial chassis able to produce antibiotics from CO2. We are developing an approach focused on the selection of the best Streptomyces strains able to produce antibiotics in a poor environment. In parallel, we are developing rich GC biobricks to implant the genes encoding the two enzymes necessary for the implementation of the Calvin cycle in these strain, namely RuBisCO and phosphoribulokinase. We are also cloning RuBisCO and PRK from a Streptomyces in E. coli to verify the activity of these proteins. We performed HPLC analysis (to test RuBisCO activity), antibiotic bioassays and growth curves in CO2 enriched atmospheres (generated by gas packs in jars). Our project is thus in line with artificial autotrophy, the first step towards carbon neutrality.
Concerning iGEM's rules and policy
We did not do any prohibited activities. We have not used any organism from the risk group. We have not released or deployed any genetically modified organism outside of the laboratory and we have not tested our product on any human (including ourselves). All our work is covered by the white list. We work with Streptomyces (S. albidoflavus, S. bottropensis, S. coelicolor, S. lividans, S. rimosus, S. ambofaciens, S. venezuelae), under safety hood and risk group 1 microorganisms, other than spore-forming bacteria (e.g. E. coli DH5-alpha, BW25113 and BL21 strains). The modifications that have been made on these microorganisms did not change the grade of these risks. We did not do research on human subjects.
About our laboratory
We were in a standard microbiology laboratory (L1) within the I2BC at CNRS. All the necessary equipment for the safety of our experiments is present within the establishment. We have open benches, chemical hoods, laminar flow hoods and biological safety cabinets. We therefore have the main materials necessary for safe handling. We did not use chemicals or dangerous organisms in our experiments, except ethidium bromide (for analyses on gel) that was manipulated under safety conditions. Our products were mainly biological materials. Autonomous environmental propagation of one or more of our organisms is possible. They may be able to develop and help the system to regulate the minimal CO2 rate. To avoid any contamination, we always handled Streptomyces and E. coli (risk 1 bacteria) under a safety cabinet. Chemical and biological waste was disposed of in dedicated garbage cans and each of our handling followed the safety guidelines of our laboratory. The laboratory's safety officers assisted us in risk management. In addition, each member of iGEM had received safety/security training or reminders regarding: Laboratory access and rules; Responsible persons (e.g., laboratory or department specialist or institutional biosafety officer); Differences between biosafety levels; Biological safety equipment (e.g., biological safety cabinets); Good microbial technique; Disinfection and sterilization; Emergency procedures; Physical biosafety; Chemical, fire and electrical safety. To manage the risk of our project biosafety/biosecurity measures were taken into account: Accident reporting (system to record all laboratory accidents); Personal protective equipment / PPE; Physical access controls; Data access controls; Lone worker or off-duty policy.