Phylogenies_of_the_mouse_blood_forming_system__TGS_
This project seeks to understand clonal relatedness between haematopoietic stem cells (HSC) and their total contribution to haematopoietic output by exploring single cells derived from different geographical locations of bone marrow niches in the mouse. Utilising naturally acquired mutations as barcodes, we will be able to track the progeny of HSCs in addition to trace their life history, from adulthood back to embryogenesis, which will enable a clearer understanding of HSC clonal dynamics. Through deep sequencing of whole genomes from HSC-derived colonies, we will be able to construct the phylogenies of stem cells isolated from different niche locations, and at the same time we will understand the dynamics of blood cell production. For phylogenetic tree building, between 60-70 single-cell derived colonies will be profiled through Whole Genome Sequencing (WGS), and a targeted bait sequencing panel will be designed after the selection of branch-defining mutations identified on the tree. Peripheral blood (PB) mature cell populations including Granulocytes, B and T cells will be profiled through the targeted bait panel, so that they can be assigned to specific branches on the phylogenetic tree and thus trace back their ancestry.