Seeing is believing – Studying chromosomally-derived circular DNA in human stem cells: Are they transcribed?
Navn på bevillingshaver
Henrik Devitt Møller
Beløb
DKK 295,000
År
2018
Bevillingstype
Internationalisation Fellowships
Hvad?
The nuclear human genome consists of discrete linear chromosomes. However, extrachromosomal circular DNA elements (eccDNAs) can mutate out of the linear chromosomes and stay inside a cell for unknown periods of time. EccDNAs carrying proto-oncogenes are found to correlate with oncogenesis, likely through the accumulation of eccDNA copies within single cells. I will develop and exploit novel tools that will allow me to track specific eccDNAs inside single human stem-like cells. I intend to answer whether generation of eccDNA leads to accumulation in dividing cells, if they are transcribed and whether eccDNAs have cellular consequences on e.g. cell morphology, cell-cycle duration, and cell viability.
Hvorfor?
There is an unmet need for sensitive in situ techniques to monitor eccDNAs role inside cells, in order to learn more about their influence on the cell phenotype, their localisation and prevalence over time. This is important as certain eccDNAs correlate with cancer progression, and eccDNA accumulation could be linked to ageing of cells. I will explore specific eccDNAs in human stem-like cells. Human stem cells are already used for medical treatments, and these cells are important for organ homeostasis as the cell steady-state depends on stem cell proliferation. Thus, potential eccDNA accumulation in stem-like cell types may have undesirable, phenotypically consequences.
Hvordan?
The research will build upon the innovative research from Zürich (in vivo imaging in single cells), Copenhagen (eccDNA detection) and Aarhus (CRISPR technology), bringing leading research fields together for substantial synergistic effects that will facilitate state-of-the-art research on eccDNA inside living human stem-like cells. Tracking tools will include the well-established LacO/LacI sensor system, available in ETH Zurich, and development of a new, more sensitive tool for DNA imaging, using the RNA-Guided CRISPR-Cas9 system.