Gene expression in microgels: The effect cytosol crowding and cytoplasm organization on cellular function

• Applicant: Professor Dr. Julian Thiele
• Subject Area: Biological and Biomimetic Chemistry
• Term: from 2015 to 2021
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 275401251

Commercial biological assays are generally performed in a state of dilute bulk solution. As a result, they do not reflect key properties of a cell including cytosol crowding and spatial organization on microscopic scales. Therefore, insights on underlying reaction-diffusion processes as well as in vivo reaction kinetics remain vague. This proposal is directed towards the development of an artificial microscopic cytoplasm to overcome these limitations. As a major cellular function, gene expression will be studied via confocal microscopy inside microfluidically prepared hydrogel particles that will serve as artificial cytoplasm with tailored porosity, heterogeneity, and functionalization. In contrast to the inherent complexity of living cells, the proposed experimental platform will provide unbiased information on the individual contribution of cytosol crowding, cytoplasm heterogeneity as well as spatial organization of reactants to transcription and translation kinetics. By manipulating relative location and diffusion rates of mRNA, DNA, polymerase and ribosomes through engineering the hydrogel complexity in a bottom-up approach, direct structure-function relationships will be derived and transferred to the situation in vivo. The anticipated results will additionally serve as a basis to precisely engineer biochemical reaction pathways by means of macromolecular and colloid chemistry and ultimately outperform conventional biological assays.

• DFG Programme: Research Grants
• International Connection: Netherlands
• Cooperation Partner: Dr. Hans Heus