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Single-cell temporal tracking of epigenetic DNA marks

Periodic Reporting for period 3 - EpiTrack (Single-cell temporal tracking of epigenetic DNA marks)

Período documentado: 2020-09-01 hasta 2022-02-28

Epigenetics, the study of the changes in organisms caused by modification of gene expression rather than alteration of the genetic code itself, has taken centre stage in our understanding of gene regulation, cellular differentiation and human disease. The process of adding a methyl group to a DNA molecule (DNA methylation) is one of the most prevalent epigenetic modifications in mammals whereby three major DNA methylation enzymes (DNMT1, DNMT3a and DNMT3b) deposit tens of millions of these "diacritic marks" in the genome creating cell type-specific methylation patterns. Although DNA methylation has been extensively investigated, key mechanisms of these fascinating events remain obscure. The project aims to engineer an advanced cellular system (named liveTAG) that will give a comprehensive view of the roles that the three DNA methyltransferases play in establishing cell-specific genomic methylation patterns and how they govern cell variability and ability to differentiate during mammalian development. The study aims to deepen our understanding of epigenetic regulation in mammalian cells which has important implications in human development and disease.
Using CRISPR-CAS genome editing technology and new knowledge gained during the project we have successfully redesigned the DNMT1 methylase inside mouse cells for selective deposition of trackable chemical entities in DNA. In parallel, we have advanced our development of high resolution analytical tools for mapping the deposited chemical entities on the genome scale in single cells.
The developed unique tools pave the way to selective chemical tracking of key epigenetic events underlying reprogramming of stem cells to other cell types during mammalian differentiation. These studies will provide new insights into the epigenetic regulation in mammalian cells which has important implications for our understanding human development and disease.
Strategy for small molecule-dependent chemical tagging of cognate methylation sites (liveTAG)