Our project will be engineering Pseudomonas putida TA7-EG to synthesise sandalwood oil components, santalol and santalene. To achieve this, we’ve incorporated several genes obtained from the Santalum album. None of these genes are known to have any virulent factors.

We chose suitable pSEVA backbones of low copy number to avoid burdening our bacteria and attain stable gene expression within our chassis.
Santalum album oil has been used to make small quantities of traditional medicines and personal care products. Santalum album oil has been previously tested on humans, and it has not caused any irritation or discomfort. In the case of oil from Western Australia (Santalum spicatum) or Hawaiian sandalwood (Santalum paniculatum), significant amounts of farnesol, which is known to cause allergic reactions, are present. However, Santalum album oil does not contain farnesol; hence, it does not cause any allergic reaction and is a safer option ^[3].

We followed all the instructions on properly using and handling common reagents used in procedures such as gel electrophoresis, Western Blotting, Coomassie, SDS-PAGE, etc.
Antibiotics were used for selection procedures. We used ampicillin, spectinomycin, streptomycin, gentamicin, kanamycin, and anhydrotetracycline and the safety hazards regarding each reagent were verified using the National Center for Biotechnology Information. Ampicillin, gentamicin, kanamycin, and anhydrotetracycline were already prepared at appropriate concentrations provided by our supervisors, whereas spectinomycin and streptomycin were obtained as powders. The powders were weighed and solubilized. All antibiotics were handled with proper care, and special instructions given by our lab instructor were followed.

As part of our project, we conducted several interviews, surveys and competitions to spread awareness and gain more insight regarding the project. As we had to interact with various groups of people, we had to ensure that all relevant laws and regulations were strictly followed. Permission from legal authorities was acquired before conducting any activities involving the public.

According to theb Regulations and Guidelines on Biosafety of Recombinant DNA Research & Biocontainment, our experiments fall under Category II, and hence, prior authorisation from the Institutional Biosafety Committee (IBSC) was given before the commencement of the experiments. We filled out all necessary applications required for the authorisation to handle GMOs and our work was approved by the Review Committee on Genetic Manipulation (RCGM) under the Department of Biotechnology.

Multiple surveys were released by the Human Practices, Outreach and Proposed Implementation sub-teams. Prior consent was sought from the participants at the beginning of the forms, and they were clearly informed that all data collected would only be used for the purpose of the research.

Prior consent to use any video recordings and images of webinars and interviews conducted as part of our project was given. A consent form regarding the use of any necessary content on online platforms was signed and given to us by the participants.

Since the beginning of PETal we ensured that we followed safe practices in all aspects of our project by identifying all the potential risks we may face in the future, from synthesising our product within the laboratory to manufacturing it in bioreactors for making consumer-based products. We talked to relevant producers and start-up companies and studied relevant literature regarding the same. More information regarding future safety measures is explained in the Proposed Implementation page.