Team:EPFL/Engagement
Education and Public Engagement
Aptasense, our biosensor that targets proteins, is based on a cell-free platform.
During our summer time working in the lab, we thought about how to integrate this ready to use, safe and overall highly modular protein expression chassis into classrooms to make the learning process more practical, fun and appealing to high schoolers.
The idea was born when we witnessed the enthusiasm of a class of high schoolers, we hosted in our lab in the beginning of July. In fact, we welcomed 25 students from the Lerbermatt gymnasium in Bern, and showed how fun and interesting learning about protein expression could be.
We showed them how to work with chromoproteins after we have explained what they are and how they are useful in synthetic biology. We were greatly surprised to witness the creativity that this activity brought in the class as showcased by this chromoprotein drawing below.
We were also interested in showing them how protein expression worked, especially considering how convenient and easy the experiments can be using our cell lysates.
We prepared a list of experiments to showcase the detection of beta-galactosidase using a colorimetric substrate (Chlorophenol red-β-D-galactopyranoside). One experiment involved adding the substrate to a cell lysate already containing the protein (BL21(DE3)) which makes the substrate turn to purple directly, proving the protein's presence. Another experiment entailed witnessing the gene translation by adding an E.Coli lacZ DNA template to a M15 lysate and DH5alpha lysate which have an alpha deletion, so the beta-galactosidase is not functional and the substrate stays yellow upon its addition the lysate reaction. But after 1 hour of incubation at 37°C we see a color change in the lysate that turns to purple, proving the presence of beta-galactosidase which was synthetized because of the transcription and translation of the lacZ gene in those lysates.
We quickly saw that explaining the processes of gene synthesis, transcription and translation was easier using the ludic set up and quick results that our cell-free expression system brings.
Growing more and more confident regarding the range of theoretical processes that we could explain in more practical ways using our cell-free, we could not miss the opportunity to attend the Open Plant forum in Cambridge which had many important figures of cell-free biology such as James Swartz and Keith Pardee in their list of speakers.
There, through participating in a variety of workshops we participated in the idea of our Educational Cell-free Mini Kit was born: a small toolbox that would enable teachers to give a live demonstration of various biomolecular processes. An easy to handle, high-impact collection of experiments that poses no biohazard as it does not contain living cells, seemed the perfect solution for explaining the concepts listed before to high school kids. A creative and novel way to stimulate each student's learning process.
Details on the Cell-Free Mini-Kit
The Cell-Free Mini Kit includes the material and experimental setup for the following four questions :
- Presence of the ß-galactosidase enzyme in a cell used for lysate production
- Expression of protein in a cell-free extract
- Interest of energy supply for biochemical reactions
- Polymerase operating mode
- Repression of a gene
The experiments will include the following procedures :
- Detection of ß-galactosidase enzyme presence in a lysate made of cell culture by colorimetric assay
- Study of cell-free in vitro expression of ß-galactosidase and kinetics of ß-galactosidase activity
- Showing the importance of an energy source by performing cell-free experiments with and without energy solution
- Testing different polymerases in cell-free environment with different promoters
- Study the principle of gene repression using a riboswitch which can be expressed in the presence of a trigger
Components for the abovementioned reactions include :
- E.Coli cell lysate of BL21, DH5alpha and M15 strains
- Energy solution, a mixture containing amino acids, salts, reducing agents
- Buffer A, contains Tris, magnesium and potassium glutamate
- Substrate, chlorophenol Red-β-D-galactopyranoside
- Linear DNA templates to be expressed in vitro (for example LacZ)
- Our toehold/trigger construct, which is single and double stranded DNA templates
The kit exists in two versions: either all the materials are freeze-dried and delivered in tubes containing the separate freeze-dried components, or be distributed in their fluid form using dry ice.
Visit and feedback from high school teachers
In October, two high school teachers from the Collège Calvin in Geneva came to EPFL to meet with two iGEM EPFL members and discuss a possible integration of the Cell-Free Educational Mini Kit in their high school's biology curriculum. We gave them a live demonstration of the experiments contained in the Mini Kit and discussed the impact it would have on the high school student's courses. The conversation was very interesting as we were until that point not sure how well the high school labs were equipped - would they have micro-pipets, -80°C freezers, incubators? Depending on their answers we would have to adapt the Mini Kit. The teachers were glad to respond to all our questions and were also very interested and already gave us a first feedback: instead of working with freeze-dried components, they would have the capacity to handle the components in their fluid form as they had micro-pipets and access to a -80°C freezer.
"Votre projet me parait passionnant et je serais très heureux de parler avec vous des implications et applications dans les classes. Il pourrait faire l'objet d'une publication dans le cadre du projet Bio-Tremplins de la faculté des sciences et du DIP Genève qui s'adresse aux enseignants et touche près de 400 personnes à travers la francophonie."
"Your project seems fascinating to me and I would be happy to talk to you about implications and applications in class rooms. It could be part of a publication in line with the project Bio-Tremplins of the science faculty of the university of Geneva that is addressed to teachers and will touch up to 400 people all over the french speaking part of Switzerland. »
« Je parle chaque année du projet iGEM à mes élèves de 4OS, alors si pour une fois il y a moyen de faire plus que de regarder une vidéo, je serai ravi de le faire. »
« Every year I talk about the iGEM project to my last year high schools students, hence if, for once, it is possible to do more than watching a video, I would be happy to do it. »
« Quel projet captivant et motivant sur lequel vous travaillez ! Il est toujours sympathique d’avoir des nouvelles d’anciens élèves et d’être informés de ce qui se fait dans les laboratoires. Nous sommes intéressés par votre kit."
« What a captivating project and motivating on which you are working ! It is always nice to hear from former students and being informed about what is being done in laboratories. We are interested by your kit. »
A sustainable solution for high schools
The next issue we had to settle was how to supply the schools with the components needed for the Mini Kit, in a sustainable way that will last longer than just the duration of this year's iGEM project. It was clear that after November 2017, our team would not have access to the lab spaces anymore and we would therefore not be able to deliver components of the Cell-Free Mini Kit ourselves. We looked into more details for the following two solutions:
- Having one of the labs at EPFL produce the lysate and energy solution; for example, as suggested by our supervisors themselves, within the scope of the laboratory initiation course for second year students at the faculty of Life Sciences, which would have made our project worth of a whole set of in-lab practical sessions at EPFL
- Contacting the company responsible for delivering laboratory material to the high schools directly
It was the second point that caught our attention after the meeting with the teachers. They gave us the contacts of BiOutils (french for bio-tools), whose mission it is to support the teaching of modern biology at local high schools and supply them with the material and competences needed. We have established a collaboration with BiOutils: we are at the moment in discussion with them and exchanging the protocols for lysate production and energy solution, to see when and how it would be possible for them to produce all the components of the Cell-Free Mini Kit on a larger scale and supply the high schools directly once the iGEM competition will be over. This solution is definitely the most satisfying as the Mini Kit will enter a list of supported cantonal experiments for high schools and biology teachers in all of Geneva would be able to order it and use it in their class.
We are very glad to have found such broad support and interest in the idea of our Cell-Free Educational Mini Kit and continue to work in the hope that it will become permanently integrated in local high school curricula. With the help of BiOutils, the Mini Kit will be produced on a larger scale and will be accessible for every high school in the region. Its future use will be ensured after the Jamboree and will, so do we hope, create a long lasting impact on education by generating infatuation for little known science fields amongst high school students in addition to bringing cell-free biology into swiss educational systems.
Feedback
Our interactions with the high school teachers were very valuable for us to gather opinions and reactions to the Mini Kit we wanted to propose. After exchanging the protocols with the teachers from Collège Calvin, they suggested to us that lyophilisation of the components would not be needed, as they were equipped to directly manipulate the solutions. They told us that it would be better for the course since the students would be the ones pipetting the solutions and preparing the mixtures and that it would make the Mini Kit more interactive. We of course agree with this statement and went on to modify our Cell-Free Educational Mini Kit protocols to include manipulation of the solutions directly and describe which amounts of reagents would be needed for delivery. Hence, after this discussion, we propose now two different kits: one lyophilised, for easy storage for low ressources high schools, and one non lyophilised, for high schools that could have access to pipettes and adequate storage for the kit. We managed to include many other aspects of their feedback into our Minikit, although two of them persist until now: The central questions of each experiment might be a bit too easy for high school students and it would be great to have two versions of the instructions, one for the students and one for the teacher.
After the visit, the two teachers told us they were very eager to try out a test-kit such as to integrate the kit before the current school high school semester ends in February. We promised to provide them with five test kits as soon as we come back from the Giant Jamboree in Boston so that they could get started with experiments!
The company BiOutils wanted to know how the individual components could be synthesized or produced, as they would taking over this part for the high schools. We quickly provided them with our detailed protocols on how to make lysate, which cell lines are optimal for cell-free protein expression, how to prepare the energy solution and which substrates and buffers are needed for the reactions.
After an hour-long conversation and several e-mails exchanged with the École Moserin Geneva, we had established that École Moser would like to include the Cell-Free Mini Kit in their curriculum but could only do so once the next academic year begins (starting Summer 2018). They are also being supplied by BiOutils and asked us if it were possible to expand the Mini Kit with certain experiments for more advanced biology courses. This suggestion was very much welcomed by us and we set out to write a manual that may serve as a roadmap for generating new Mini Kit experiments. At the moment we are in discussion with them on which experiments they would find useful and how to realize those with the Mini Kit.
