Single molecule observation and manipulation of gene expression dynamics to dissect mechanisms of cell cycle entry

Project Information

RNAREG

Grant agreement ID: 677936

DOI

10.3030/677936

Project closed

EC signature date 30 November 2015

Start date 1 April 2016

End date 31 August 2021

Funded under

• EXCELLENT SCIENCE - European Research Council (ERC)

Total cost

€ 1 500 000,00

EU contribution

€ 1 500 000,00

Coordinated by

KONINKLIJKE NEDERLANDSE AKADEMIE VAN WETENSCHAPPEN - KNAW
Netherlands

Periodic Reporting for period 4 - RNAREG (Single molecule observation and manipulation of gene expression dynamics to dissect mechanisms of cell cycle entry)

Reporting period: 2020-10-01 to 2021-08-31

Summary of the context and overall objectives of the project

Cell division is central to live as a multi-cellular organism. Every human develops from a single cell to a full grown person containing billions of cells through a series of many cell divisions. However, cell division must also be controlled precisely, as excessive cell division might lead to cancer. The ability of a cell to undergo cell division is dependent to a large extent on precise expression control of its genes. Certain genes must turn on, while other must turn off. In this project we develop novel technologies to understand how genes are turned on and off and we apply these technologies to understand gene expression control during the cell cycle.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

So far in this project, we have developed new technologies to study gene expression control. Specifically, we have developed a method to visualize gene expression in single living cells using high resolution microscopy, and we have developed a second technique to turn genes on and off at will, using high throughput CRISPR technology. We have applied these technologies to study gene expression changes during the cell cycle, to determine when genes are turned on, on what the molecular mechanisms are that drive these gene expression changes. Furthermore, we have characterized changes in gene expression that occur during the cell cycle, and have identified a set of several hundred genes that show a new type of regulation. Finally, we have modified these methods to allow visualization of viral gene expression as well.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

We have made several contributions to the research field that go beyond the state of the art. First, we have observed mRNA translation (the final step of gene expression) in real time for the first time in living cells. Second, we have performed quantitative analysis of gene expression changes during the cell cycle with unprecedented temporal resolution. Third, we have developed new methods to turn every gene in the genome on or off to characterize the effects of gene expression changes on for example cell cycle progression or cancer drug sensitivity. Finally, we have developed tools to visualize viral infection with single-molecule sensitivity in living cells for the first time. We envision that applying these technologies to a variety of different genes will provide a detailed mechanistic understanding of gene expression control during the cell cycle, as well as in other processes.

Human cancer cell in which translating mRNAs are visible.