Team:ULaVerne Collab/Collaborations
Collaboration with the Stanford Team:
Collaboration with the Stanford Team:
For our collaboration with the Stanford Team, we mainly talked to Alec Lourenço who was an alumni from a high school iGEM team that did a collaboration with the University of La Verne back in 2015. Now for this year, he had contacted our advisor for a chance to collaborate with us. As part of the Stanford team, they have been working on an environmental project to try to accelerate the time it takes for evolution. They are using a cell-free system since some advantages of using one are such there is a faster synthesis rate, and due to no cells being present, there is a higher tolerance to the toxic products. Using their experiment materials, they offered to be able to run our constructs for us if we had sent them some of our sequences.
Our meeting with Alec from team Stanford. He was part of the first team in ULV and wanted to collab. He introduced us to what cell-free is and how to use it.
Team members, Seth Barrington and Cynthia Basulto, working hard in lab on the Standford collaboration. While other members were setting up the plate reader, Seth and Cynthia prepared each well to be read on the 392 well plate. The data that we obtained from our plate reader was then shared back with the Standford team. After analyzing the data, we both got inconclusive data since both data sets were fluctuating.
Collaboration with the Australian DISCO Team:
When we first began our project, we were referred to the Australian Team by our advisor when the student leads began their project during their molecular class. For this collaboration, we talked to Alex Kelly who was one of the project leaders of the DISCO Team. We learned that when making the single-chain insulin (SCI) constructs they had modified or added the linker, AA substitutions, fusion protein tags, pI, while avoiding the patents of the commercial insulin, which helped point our construct design to the proper direction. He also pointed out to us that we do not need to make the insulin protein thermostable, as it is already stable during transportation. He is currently working on this project today, and encourages us to help him troubleshoot it too. He also introduced to us Open Insulin, a community aiming to manufacture Open Insulin, which his team collaborated with, and stressed to us about how important it is to make it open source so that big companies will not be able to monopolize the insulin industry like right now.
Collaboration with the Open Insulin Group:
For our collaboration with the open insulin group, Alex Kelly from the Austrailian DISCO team had actually referred us to the Open Insulin group from Counter Culture Labs since they were already working together on Insulin before we started our project. With this group of people, we have been in contact with them through video calls so we can tackle topics that we might not know about or to troubleshoot some experimental areas we were having trouble with. Due to our common cause in trying to synthesis insulin, sharing data in regards to using ecoli when they use yeast aids in collecting more data.
Our first meeting with the Open Insulin group and introducing ourselves. This meeting was just to give everyone an idea of what we were working on and possible collaborations could possibly happen.
The Open Insulin group would have weekly meetings on Wednesdays which the group would review important points which they needed to talk about. They would also have different experts attending the video conference and would give their opinion. We would also update them on how our results were going as well as share protocols.
Collaboration with Yann from the Open Insulin Group:
From the Open Insulin Group, we were able to talk to Yann about our project. He was able to give us some suggestions on our protocol for extraction methods such as removing EDTA from our osmatic shock buffer to aid in our protein purification. We were able to discuss our project in detail over a video call since we couldn't in the weekly meetings. He was able to give us suggestions on how we could potentially better our protein expression since we were having difficulties. He suggested that we should consider adding a GGS linker. We were able to ask information about the manufacturing side of Insulin and he agreed with us using e-coli as a chasi. He also explain for our small scale project protein purification using SDS-page was the best route to take. Since other manufacturing companies do one chain at a time then put them together
Team members Cynthia Basulto, Karen De Leon, Catherosette Meas, and Magaly Aguirre were able to talk to Yann to troubleshoot some parts of our SDS-page gels. We were also able to share our online notebook, Padlet, with him in order to stay updated with what we have been doing in the Wet lab portion.
Collaboration with the Bilkent-UNAMBG iGEM Team:
Team Bilkent-UNAMBG planned to measure the activity of their single-chain insulin. We were able to Skype with their team and discuss strategies with each other. We discussed our projects in detail while asking questions to each other. Their advisor was also able to give us points on the characterization aspect of our project. They also suggested doing a Western plot would be more beneficial for what we are trying to achieve. One good thing to keep in mind for future direction is if there is too much protein expression in the periplasm, it could to toxic to the cell. Their advisor pointed out using a lower promoter to expression could be the best way to not stress the cells.
Their advisor also pointed out that having our HIS-tag in the middle of two big proteins could be an issue for purification purposes. We were able to discuss our reasoning of why we continue our project as well as explained why we included linkers. For future directions, Team Bilkent-UNAMBG suggested we would quantify how much protein is released from the cell for math modeling. We could potentially use Pymol to find the RSMD score. This would show the distance between two molecules.
Collaboration with the Moscow iGEM Team:
Wetlab:
1) We've made a construct of Insulin by Gibson Assembly in pSB1C3 vector. We were able to dry it on the filter paper and send it via mail to their postal address. We asked them to purify via Ni-chromatography and to also confirm with SDS-PAGE.
2) We shared with them our Padlet notes and looked through it to keep up to date with what we are doing. The map of our construct was also sent as a .dna file.
Dry lab:
1) we wanted team Moscow to perform docking and molecular dynamics with our mutants. So we send them 3 PDB files of corresponding mutants, 1 PDB file of initial insulin, 1 PDB file of Insulin receptor.
2) To make our modeling closer to what we wanted to achieve, we send a file describing the parameters: pH-value, pI-value and other desired characteristics of your protein.
Our side of the collaboration:
There was 3 different collaborations that we participated with the Moscow team. Each collaboration had different rules which Team Moscow was able to explain to us. They were also very friendly and helpful when we had a question about one of the collaboration ideas. The four collaborations are the following listed below.
Chess
Games were played with 5 min per player time control until one of the player was in the lead after an even number of played games. Even though our team lost, we were able to still have fun. Team member Magaly Aguirre was able to play against team Team Moscow and Team uOttawa.
Each group had fun in each competitive game between different groups from different places. Team Moscow was able to set an online Blitz Chess tournament with different igem teams. At the end of the day, some teams expressed that they were beginners but still wanted to be part of the tournament.
iGEM2019 Spartakiade
Being in lab can be so stressful and most of the time, everyone is sitting for multiple hours. Team Moscow decided to launch "iGEM2019 Spartakiade" where teams from all over the world can participate in easy, funny, positive exercise marathon. We just followed the rules provided by Team Moscow. The wanted to unite teams in a simple marathon to receive positive vibes and to make new friends.
These are only some of the videos that we were able to upload on our social media accounts. The whole ULaVerne_collab team can agree that it was a brilliant idea to be able to collaborate and do these videos. We didn't really notice how stressed we were until we were able to reflect on it. Doing these exercises was a great stress reliever and it was also fun to step back from our busy schedules.
Survey
Team Moscow is working on a portable biosensor that can detect the presence of Borrelia spp., which can cause Lyme disease in ticks. They created this survey to learn and to create awareness of tick-borne diseases among the public. They required a minimum of 5 participants from our team to do the survey. After our team completed the survey, we were able to let Team Moscow know of our completion.
