SRIMEM · Super-Resolution Imaging and Mapping of Epigenetic Modifications
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2018-09-01 → 2020-08-31
- EU contribution
- €171,461
- Participants
- 1
- Scheme
- MSCA-IF-EF-ST
Lines connect the coordinator with its partners.
Results in brief
Super-Resolution Imaging and Mapping of Epigenetic Modifications
Super-resolution microscopy has become a tool of choice for biologists for visualizing cellular structures. Central to any of the super-resolution methods are labeling probes that recognize and bind the target of interest to enable us to localize it using fluorescence microscopy. However, there is a great need for finding suitable binders that introduce minimal artifacts such as linkage error in the imaging data and enable multiplexed imaging while allowing quantification of target protein species. Moreover, the stoichiometry of labeling by binders is completely blind folded hitherto. In this project, we addressed various issues related to the binders to improve the imaging capacity and quantification of binders’ performance. We developed novel imaging probes that introduce minimal linkage error. We also introduce a DNA nanotechnology-based origami platform to quantify the absolute efficiency and achievable resolution of any labeling probe. Furthermore, we developed a multiplexed super-resolution imaging approach to image unlimited numbers of targets of interest. The latter technology will be useful for high throughput imaging of cellular proteins and genome architecture as well as epigenetic state of the genome.
Data: CORDIS, © European Union
Project objective
Epigenetic marks are posttranslational modifications of chromatin that act as gene regulators. Although every cell-type contains the same DNA sequence, the epigenetic marks dictate specific function of each cell-type. Epigenetic modifications are both heritable and dynamic, and can be treated enzymatically to reverse. The dynamic marks sometimes lead to aberrant gene regulation in cells, causing diseases such as cancer, Alzheimer’s, and diabetes. Therefore, epigenetic state of individual genes can be used to identify the aberrant genes to reverse them.In this project, a novel assay for simultaneous identification of epigenetic marks and their genomic position is proposed. State-of-the-art DNA-PAINT super-resolution microscopy, developed by Prof. Jungmann, in combination with immunofluorescence in situ hybridization (iFISH) will be used to identify the epigenetic marks in human cells with very high precision (<5 nm). The novelty of this assay is that, for the first time, it will allow to read the epigenetic marks, their genomic and 3D position in the nucleus simultaneously, for precise mapping of the epigenetic state of genes in individual cells. The assay can be used as a tool to identify aberrant marks in cells for diagnosing diseases caused by these modifications. Therefore, it has a high potential for both research in biotechnology and diagnostics.
Original text from CORDIS.
Participants
- MAX-PLANCK-GESELLSCHAFT ZUR FORDERUNG DER WISSENSCHAFTEN EV · MUNCHENCoordinatorGermany
Links
Data: CORDIS, © European Union