Department Bionanoscience
Principal investigator Marileen Dogterom
E-mail address m.dogterom@tudelft.nl
Website https://dogterom-lab.nl/
Aster positioning in Droplets with Nucleus/Actin cortex
Suitable as a BEP? Yes
Suitable as a MEP? Yes
Suitable as an Academic Research Project? Yes
Techniques:
- Droplets
- Microfluidics
- Microscopy (confocal)
- Image analysis
- simulation (optional/Cytosim)
Correct positioning of the (mitotic) spindle is an essential process in cell division, as its position plays an important role in determining the division plane and hence the size of the daughter cells. Learn how the microtubule aster positions inside a 3D cell-like confinement, and how the force balance changes in response to constraints of the environment and additional components. We use in vitro reconstitution experiments of microtubule asters in confinements, where we can control the environment and aster components. Objectives – 1. encapsulate minimal asters, 2. quantify aster position, 3. additional effect of organelles like nucleus, actin, dynein and, 4. (optional) simulate aster positioning and compare with experimental results to understand the force balance.
Further reading (click to link to article)
Reconstitution of basic mitotic spindles in cell-like confinement. Roth et. al. doi: https://doi.org/10.1101/770602
Characterize ParM filaments and their force generation from the ParMRC DNA segregation system
Suitable as a BEP? No
Suitable as a MEP? Yes
Suitable as an Academic Research Project? Yes
Techniques:
- Microscopy (TIRF)
- Optical tweezers
- Image analysis
A promising candidate for minimal DNA segregation in synthetic cells are bacterial cytoskeletal systems. More specifically, we are interested in the bacterial ParMRC system, where filaments are formed by ParM and a ParRC-complex is able to bind these filament ends. When two growing ParM filaments are oriented anti-parallel, a simple spindle can be created to segregate the parC DNA at the opposite poles. In our lab, we aim to reconstitute functional minimal spindle for DNA segregation based on ParMRC and study in detail.
Objectives – 1. characterize biophysical properties of ParM in vitro, 2. measure filament growth rate and force, 3. measure the sliding rate and force of anti-parallel ParM overlaps in the spindle.
Further reading (click to link to article)
A Bipolar Spindle of Antiparallel ParM Filaments Drives Bacterial Plasmid Segregation. P. Gayathri et al. Science 2012 doi:10.1126/science.1229091