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Investigating the utility of the optogenetic toolbox for cell cycle synchronization in budding yeast

Bartelds, Mart (2019) Investigating the utility of the optogenetic toolbox for cell cycle synchronization in budding yeast. Master's Thesis / Essay, Molecular Biology and Biotechnology (2016-2019).

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Abstract

Cell division in eukaryotes is achieved via a conserved and tightly controlled protein network. In order to study processes that happen at specific stages during this division cycle it is important to have a culture with synchronized cells. Currently used synchronization methods often use the ‘arrest-and-release’ strategy, in which cells are arrested at a specific point in the cell cycle using chemicals or conditional mutants. Releasing the cells from the arresting conditions results in a synchronized re-entry to the cell cycle. However, these methods usually have severe side-effects on cell physiology and the switching between the restrictive and permissive state is slow. To overcome these limitations, optogenetic systems may be used, as these systems can offer exact molecular control over diverse cellular processes and switching between two states can be achieved rapidly. To identify potential targets for optogenetic control an overview is given of natural existing cell cycle arresting pathways. Two exiting optogenetic systems were identified that utilize these pathways. Since these systems were not designed for cell synchronization, ways to further improve these systems for cell synchronization were discussed. Moreover, two other pathways were identified that showed high potential for cell synchronization. Finally, two papers are discussed that developed systems for direct control of the expression or degradation of key regulators of the cell cycle. Although these systems can potentially invoke less severe side-effects, the arrest is less stringent. Overall, although most optogenetic-based systems would require more optimization to be able to compete with existing cell synchronization methods, these system have the potential to overcome the limitations of the current synchronization methods in the future.

Item Type: Thesis (Master's Thesis / Essay)
Supervisor name: Milias Argeitis, A.
Degree programme: Molecular Biology and Biotechnology (2016-2019)
Thesis type: Master's Thesis / Essay
Date Deposited: 01 Nov 2019
Last Modified: 04 Nov 2019 10:06
URI: https://fse.studenttheses.ub.rug.nl/id/eprint/21168

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