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Development Of A High-Throughput Technique To Identify The Mechanism Of Replicative Aging In Single Yeast Cells

Reference : PhD Gilles CHARVIN

Offer publication : April 5, 2016

Background: Finding causal relationships has been a major challenge in ageing research. Through the development of well-controlled systems, our aim is to find such relationships which will open up new avenues for ageing research and - in the long run - may unravel the causes of many age-related diseases in humans.

In the context of the complexity of organismal ageing, the cellular ageing of baker’s yeast, a unicellular eukaryote, has been a powerful model system. However, the full potential of using yeast as a model for ageing of dividing cells has not been fully unlocked. A major problem has been that, with
increased age, the old cells are surrounded by up to a billion progeny cells due to the exponential growth of the population. Only very recently, the Veenhoff and Charvin laboratories, amongst others, have developed novel experimental methods that make aged cells tractable, both on a population
and single cell level. This now allows us to address specific ageing theories, and leads the way to developing higher-throughput methods that eventually allow genome-wide studies.
Specific aims: Yeast has a unique advantage in ageing research because it divides asymmetrically: while the mother cell ages, the daughter cell is born rejuvenated. A major challenge and opportunity is therefore to determine precisely how a rejuvenated daughter yeast cell differs from the aged mother cell, as this would pinpoint the molecular players that define cellular age. In the current proposal, we focus on protein abundance and
subcellular localization, two of the critical parameters that define the protein’s functional state. Imaging single yeast cells over a lifetime by microfluidic methods, such as those developed by the Charvin lab, is uniquely suited to probing both parameters as a function of age. However, these methods are currently low-throughput. With the ultimate aim in mind to study the subcellular localization and abundance of the entire proteome during aging, the first aim of the project is to develop new higher-throughput designs of microfluidic devices and develop tools for high-content automated image analysis.

These developments are needed to exploit, for the first time, baker’s yeast to its full potential in ageing research, decades after the first reports on ageing and rejuvenation in yeast.
Complementary to these investments in method development, we will use the existing low-throughput, but still state-of-the-art, technology to investigate specific ageing theories. In particular, recent data from the Veenhoff laboratory suggest that ageing Nuclear Pore Complexes (NCP’s) – the conserved sole gateway to the nuclear interior of a cell – may be important for ageing of dividing cells. The second aim of this project is therefore to
investigate the effects of ageing NPCs in ageing yeast cells. Here, the PhD student will focus on the role of NPCs in orchestrating the localization of specific chromosome regions at the NPC.

Complementary expertise: The expertise of the Veenhoff laboratory is in yeast ageing and nuclear pore complex biology. The expertise of the Charvin laboratory is in yeast ageing, the design of (yeast ageing) microfluidic devices, and development of tools for high-content automated image analysis.

- WISHED SKILLS : Development of a high-throughput technique to identify the mechanism of replicative aging in single yeast cells

- EXPERTISES WHICH WILL BE ACQUIRED DURING THE TRAINING : Yeast genetics Microfluidics Quantitative imaging and image processing

Your application

Application Deadline : Dec. 31, 2016

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