Aim: To discuss our project with the future generation of scientists, to introduce them to synthetic biology and methods within synthetic biology, and to give them an opportunity to lab in a university lab using the previously discussed methods.

We started with trying to find a group of high school students that would like to collaborate with us. Thankfully, the previous iGEM team of Uppsala had already established contact with a high school that wanted to return this year. Additionally, our course responsible had talked to another school about iGEM and possibly collaborating. From this, we established contact with both schools and set out a preliminary schedule. We managed to visit one of the schools before their summer break, where we presented our project and talked about synthetic biology. The high school students and their teacher then went on summer break, while we started preparing the experiments and the lectures.

In total, 50 high school students from two high schools in Uppsala visited Uppsala University for a week. We started the week with a presentation about our project and our results, since this took place in the very last week of iGEM. We also held a lecture about methods in synthetic biology, starting from the central dogma of biology and moving towards more complicated subjects. Margareta Krabbe and Anthony Forster held lectures about synthetic biology and their work as well, which was greatly appreciated both by us and by the students.

After a session thoroughly discussing what we were going to do and safety measures, the students were taken to the lab. We had one school work in the morning, and the other one in the afternoon. We started with a safety tour of the lab so that the students would be prepared in case of an emergency. However, we also took the opportunity to show our 37° room and some of our equipment, which seemed to spark an interest in some of the students.

After the tour, we got to work. The students were asked to choose 3 “mystery plasmids” from a selection and got instructions on how to transform competent bacteria. The plasmids contained genes for different chromoproteins and fluorescent proteins that we constructed using sequences given to us by Letian Bao, PhD student at the Department of Cell and Molecular Biology at Uppsala University. We also made the competent cells and wrote an easy-to-understand protocol in both Swedish and English so that both students and lab leaders could understand. The lab leaders consisted of members of the Uppsala iGEM team who helped the students, oversaw safety and showed techniques such as spreading techniques. The students worked with chloramphenicol resistant bacteria, except for one strain that was kanamycin resistant but was still plated on a chloramphenicol plate. One important topic we discussed was using genes for antibiotic resistance in order to select for bacteria that had taken up plasmids. In order to show proof of concept, two groups plated the kanamycin resistant bacteria on kanamycin-plates to show the groups what would happen if the same strain was plated on plates using different antibiotics, and how that could be used in real life.

The cells were left to grow over two nights to allow for visible colour, and the students returned to have a session discussing the experiment. They looked at their plates in small groups and discussed the results with lab leaders. Understanding why the kanamycin-resistant bacteria did not grow on the chloramphenicol plates was one important aspect we wanted the students to understand. The other one was to compare the different strains of bacteria: Did the chromoprotein or the fluorescent proteins expressed affect bacterial growth? If so, why would that be? If there were any unexpected results, we also tried to discuss with them how that might have happened. They were also comparing the different strains with other groups so that they had the opportunity to see all chromoproteins in action. The classes then gathered for a final goodbye session and gave us some feedback on our work.

After the lab week, we froze remaining plasmids so that the next year's team could use them. We went through the feedback, which was overwhelmingly positive, and talked about how the sessions could have been improved.

Promoters, CDS for chromoproteins and fluorescent proteins were given to us from Letian Bao, PhD student at the Department of Cell and Molecular Biology at Uppsala University. We wanted a strong consecutive expression, so the target genes were placed after a suitable promoter, ligated into either pSB1C3 or pSB1K3 and transformed using E. coli DH5α cells [Bao L, 2020].

CDS used:

mRFP1
amilCP
amajLime
fwYellow
aeBlue
cjBlue
Promoter was J23110.

CDS and promoter were ligated into plasmids pSB1C3 or pSB1K3.