Safety

Biosecurity and biosafety are important concepts when working with genetically modified organisms(GMOs). Hence, during the wet lab work we have been always taking precaution of any possibility of release to the environment. On the other hand, assuring people security is another priority on our project. Thus, we hope to develop our idea in order to improve the farmers’ production taking into account their health.

Our project: No GMOs

The project was approached in order to avoid any type of GMO escape to the environment. According to the Peruvian law, it is forbidden to release these modified organisms to the wild due to the lack of knowledge about the impact that could have on the environment.

Since we were deciding which organism would be ideal for our project, we stated as one of our main requirements that our chassis would be classified in the first Biosecurity Level (BSL-1). Pseudoalteromonas nigrifaciens is a nonpathogenic bacteria that was first isolated from butter by White in 1940. This bacteria is considered to be innocuous for humans and would only cause injuries to sea cucumbers.

Other than that, it has been documented that depending on its strain it would develop different antibiotic resistance. Therefore, to determine the antibiotic resistance of our chassis strain, P. nigrifaciens ATCC 19375, we made a growth test overnight in a control situation and adding different antibiotics at different concentrations:

Fig1. Growth curves of P.nigrifaciens in PB medium with different antibiotics. Control: No antibiotic.

Laboratory work

Our institution

We work under the guidelines established by a biosecurity committee from the Faculty of Science and Philosophy of Cayetano Heredia Peruvian University. These entities are the Division of Research, Science and Technology (DUICT), and the Technical Office of Health and Safety at Work (OTSST). At the Laboratories of Research and Development (LID) where our laboratory is located, they make sure that investigators follow the rules and regulations of research work and count with trained personnel in laboratories, besides establishing biosecurity levels, security signage, protective equipment and residue management. These guidelines have been made in accordance with the rules of the National Institute of Health (INS) of Peru and National Center of Epidemiology, Prevention and Control of Diseases

Our actions

Our team is working in the Individual Molecules Laboratory of PhD Daniel Guerra at LID, which is classified as a laboratory BSL-1. Before experimenting, we had been taught and trained by experienced members from this laboratory about how to manage dangerous reagents such as ethidium bromide, following molecular protocols and correct biological material and equipment usage, making us able to handle the process of molecular cloning safely and securely. Besides that, the first time we would do a new experiment we would be looked after by our instructor and with all the basic biosafety and biosecurity measures.

Humans subject research

We followed all the necessary steps to register our project on the platform dedicated to monitoring the research of our university called SIDISI, so that it goes through the review of the institutional ethics committee. Thus, after approximately a month, we were able to start with the activities planned with the farming communities of the department of Junín.

Future plans

According to our goal, we want to produce a functional antifreeze protein by an expression system in P. nigrifaciens.To improve safety of our project, we want to design a hermetic bioreactor in which we could store and maintain our chassis out of contamination and only extract the AFP from it to be spread to crops. Although we are developing only the initial phase of this project on iGEM, we still want to approach other aspects that would be important for the better production and usage of this product, such as anti freezing efficiency. For it, we will conduct a pilot stage with activity tests in test tubes, thereby we will know what AFP works best and in what concentration the protection is optimal to reduce frosting.