Department Bionanoscience
Principal investigator Arjen Jakobi
E-mail address a.jakobi@tudelft.nl
Website https://cryoem.tudelft.nl
Characterization of dynamin-like proteins as possible candidates for assisting synthetic cell division
Suitable as a BEP? Yes
Suitable as a MEP? Yes
Suitable as an Academic Research Project? No
Techniques:
- Cloning
- Molecular biology
- Protein expression and purification
- (Cryo-)electron microscopy
This project focuses on biochemical and structural studies of dynamin-like proteins (DLPs) that may be suitable for use as a minimal divisome for synthetic cell division. DLPs are known for their roles in membrane remodeling processes, including membrane fission. The project will focus on DLPs that possess specialised targeting signals as potential divisome candidates. These are interesting candidates in the context of synthetic cell division due to their capability to promote membrane scission in reverse topology. We will first establish a construct of one of the candidates through cloning and produce it with protein purification techniques. Then, biochemical characteristics of the protein will be investigated using mass photometry. Structural information will be obtained through negative stain and/or cryo-electron microscopy.
Further reading (click to link to article)
https://pubmed.ncbi.nlm.nih.gov/37798563/
Design and fabrication of MEMS-based sample carriers for time-resolved electron cryo-tomogrpahy
Suitable as a BEP? Yes
Suitable as a MEP? Yes
Suitable as an Academic Research Project? No
Techniques:
- Nanofabrication
- Simulation
- Fluorescence microscopy cryo-electron tomography
Achieving reliable and reproducible time-resolved cryo-electron microscopy (TR cryo-EM) requires careful attention to multiple aspects of the experimental workflow. This is essential for capturing and resolving transient intermediate states of macromolecules as they undergo chemical reactions. To this end, systematic refinement of individual procedures, such as plunge freezing, sample carrier design, and the physical understanding of each step, is critical.
Currently, we are developing a single-membrane silicon nitride (SiNx) sample carrier for imaging neurons, which represents an initial step toward the fabrication of electron-transparent nanochannels for TREM. In this project, contributions to the characterization of SiNx membranes using TEM, cryo-CLEM, or confocal microscopy, as well as participation in sample preparation experiments with the fabricated chips, would be highly valuable.
Further reading (click to link to article)
https://doi.org/10.1002/adfm.202313705