Supporters
Our team accomplished great things in the project Chlamylicious over the course of this year’s iGEM competition: We adapted a toolkit for the green algae C. reinhardtii with parts for successful and easy cloning of any transcription unit, which we implemented for the expression of the PET-degrading enzyme PETase by the algae and designed a bioreactor for its cultivation under constant monitoring of all parameters. These were our greatest supporters:
- our host laboratories Experimental Biophysics and Molecular Genetics from the Humboldt University of Berlin, who shared their infrastructure with us
- our supervisors Prof. Dr. Schmitz-Linneweber and Prof. Dr. Hegemann
- the principal investigator Simon Kelterborn, who guided us with his expertise in transformations of Chlamydomonas and CRISPR-aided genetic engineering
- the biological technical assistants Heide (Biophysics lab), Conny, Thea and Irina (all Genetics lab), who enabled us to launch the wet-lab
- computer scientist Vincent Will, who implemented the design for our website
- Prof. Dr. Ringrose, who coached us on how to create a good scientific poster
- electrical engineer Markus Schulz, who helped designing and wiring the electronics for our bioreactor
- technical engineering student Alejandro Gómez Jimenez from the Technical University of Berlin, who wrote the CAD-models for all parts of our bioreactor
- the Interdisciplinary Workshop „Matters of Activity“ of Humboldt University, who laser-cut our hardware
- the Mechanical Workshop of the Physics Department of our university
Re-used parts from different sources
| Part name | Source |
|---|---|
| AR | Plasmid p114, Hegemann lab (Biophysics Department, Humboldt University of Berlin) |
| ARS | Ordered oligos from IDTSequence:https://www.uniprot.org/blast/?about=A8ISJ6[1-26]&key=Signal%20peptide |
| AmpOri | Amplified from MoClo plasmid pICH47732 |
| Ble/Scp: | Ordered oligos from IDT |
| GLE | Uniprot Q93WN9[1 - 28] |
| 3xHA-Tag | Ordered oligos from IDT |
| His-Tag | Ordered oligos from IDT |
| Hyg | Amplified from p360, Hegemann lab (Biophysics Department, Humboldt University of Berlin) |
| MHETase (wildtype) | 2019 iGEM team University of Kaiserslautern, (C. reinhardtii) |
| MHETase (wildtype II) | Weber lab (Molecular Structural Biology, University of Greifswald), (E. coli) |
| MHETase (Mutant W397A) | Weber lab (Molecular Structural Biology, University of Greifswald), (E. coli) |
| Paro | Amplified from p114, Hegemann lab (Biophysics Department, Humboldt University of Berlin) |
| PETase | Ordered oligos from IDT, Codon usage is for C. reinhardtii |
| PETase (E. coli) | Adgene's plasmid: plasmid #112203 |
| PtxD | Sequence: AF061070.1Ordered from GeneCust |
| PsaD | Plasmid 51, Hegemann lab (Biophysics Department, Humboldt University of Berlin) |
| PsadIntron | Self designed, intron designed after those of the AR-Promoter |
| RbcS2 | Amplified from p141 pCrP2-SaCas9 (RBCS2, G), p016 |
| RFP | p42, Hegemann lab (Biophysics Department, Humboldt University of Berlin) |
| Spec | Amplified from an Addgene MoClo Plasmid pAGM9121 |
| YFP | Amplified from YFP Sequenz of p135 pCr3-Ble2-YFP-Sp-APH plasmid >pAGM9121Overhang PCR: Amplification |
Timeline and responsibilities
As the first team from the Humboldt University ever to form and participate in the iGEM competition we had to organize the following to establish C. reinhardtii as a chassis for synthetic biology in the iGEM community:
- Find a professor and department able to supervise the iGEM project. As there is no course of synthetic biology taught at our university we referred to the Molecular Genetics and Biophysics lab groups.
- Found a team of students, who were able to conduct the project research during their studies without earning credits for it. Our university so far does not offer any courses for the participation of students as an iGEM team. By June 2018 the main core of our iGEM team was brought together.
- Brainstorm for interesting and viable SynBio projects, which took place in July 2018. Most of us had little or no prior knowledge in synthetic biology.
- Research more on the possibilities of C. reinhardtii as a SynBio tool and familiarize ourselves with project management conducts from July-October 2018.
- Start fundraising initiatives in July 2018.
- Get part of the team started with working in the wet lab in November 2018.
- Organize Human Practice events and possibilities of outreach to the broader public from November 2018 - October 2019.
- Get a second lab team to finish the wet lab work from April-October 2019.