Project description DEENESFRITPL Advanced study into the mechanisms of transcriptional memory Transcriptional memory is a phenomenon during which cells primed with particular stimuli show increased gene expression after re-stimulation at a later time. Such memory of gene expression states is essential for the development and maintenance of tissues. Funded by the Marie Skłodowska-Curie Actions programme, the IMMUNE-GENEMEMO project is based on the discovery of genes with a strong transcriptional memory of prior interferon-gamma (IFNγ) activation, creating a larger proportion of cells expressing the target gene. The project aims to identify novel transcriptional memory maintenance factors in human cells using an efficient high-throughput cell sorting strategy combined with genome-wide CRISPR-Cas9 mutagenesis protocols, and establish the role of these components in the stability of dendritic cell identity. Show the project objective Hide the project objective Objective Memory of gene expression states is essential for development and maintenance of tissues. Chromatin, a protein DNA complex, is implicated in the process. Local, chromatin feedback loops are involved in maintenance of gene silencing, but analogous mechanisms for preservation of active states are unknown. In order to reveal feedback loops for active gene expression states it is crucial to uncouple them from ongoing transcription, as it occurs in the case of maintenance of silent states. Such partition takes place during trained immunity – memory of the innate immune system. A key paradigm is interferon gamma (IFNγ) stimulation. While IFNγ induces many genes, a subset of those is maintained in a poised, inactive state that allows for rapid re-activation at a later time – an event called transcriptional memory.In my ongoing research I have discovered novel genes that show strong transcriptional memory of prior IFNγ activation and have obtained initial mechanistic insights into the role of chromatin in the process. Importantly, I discovered that memory results in a larger proportion of cells expressing the target gene. This forms the basis of a cell sorting, high throughput assay that I have developed. I am now in a unique position to capitalize on those discoveries and tools. I aim to identify novel transcriptional memory maintenance factors in human cells using an unbiased approach based on an efficient cell selection strategy combined with genome wide, CRISPR-Cas9 mutagenesis protocols. Secondly I wish to establish the function of the identified components in the stability of dendritic cells identity after differentiation form monocytes.The multidisciplinary and unique perspective of this project: combining gene expression analysis and selection strategies with immunological expertise will advance our understanding of the mechanisms underlying maintenance of gene expression states and contribute to the development of new immune therapies and vaccines. Fields of science natural sciencesbiological sciencesmolecular biologymolecular geneticsnatural sciencesbiological sciencesbiochemistrybiomoleculesmedical and health sciencesbasic medicineimmunology Keywords transcriptional memory trained immunity IFNγ signalling CRISPR-Cas9 genetic screen fluorescence-activated cell sorting degron system monocyte dendritic cell Programme(s) H2020-EU.1.3. - EXCELLENT SCIENCE - Marie Skłodowska-Curie Actions Main Programme H2020-EU.1.3.2. - Nurturing excellence by means of cross-border and cross-sector mobility Topic(s) MSCA-IF-2020 - Individual Fellowships Call for proposal H2020-MSCA-IF-2020 See other projects for this call Funding Scheme MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF) Coordinator UNIWERSYTET GDANSKI Net EU contribution € 252 078,72 Address Ul. bazynskiego 8 80309 Gdansk Poland See on map Region Makroregion północny Pomorskie Trójmiejski Activity type Higher or Secondary Education Establishments Links Contact the organisation Opens in new window Website Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Other funding € 0,00 Partners (1) Sort alphabetically Sort by Net EU contribution Expand all Collapse all Partner Partner organisations contribute to the implementation of the action, but do not sign the Grant Agreement. THE GENERAL HOSPITAL CORPORATION United States Net EU contribution € 0,00 Address Fruit street 55 02114 Boston ma See on map Links Contact the organisation Opens in new window Participation in EU R&I programmes Opens in new window HORIZON collaboration network Opens in new window Other funding € 177 265,92
