Lineage- and stage-specific gene expression in human hemopoietic cells

grantor: University of TorontoThis study describes the establishment of a system for mapping lineage- and stage-specific gene expression in human hemopoietic precursors, and demonstrates its ability to generate information relevant to mechanisms that control proliferation and differentiation during hemopoietic development. Two approaches were used to define patterns of gene expression at specific stages in hemopoietic differentiation. The first of these is global RT-PCR which amplifies all polyA$\sp+$ mRNA from a single cell. Hemopoietic precursors in different commitment states are phenotypically similar and cannot be separated from one another or purified to homogeneity using existing methods. The ability to study gene expression at the level of a single cell can provide an alternative approach to achieving homogeneity. The second strategy is sibling analysis, which identifies retrospectively the developmental potential of a cell subjected to PCR by analysis of the fate of its siblings. I was able to collect more than 200 cDNA samples from cells of different lineages and different stages of development. The set was then used to determine expression patterns of a wide panel of genes (RBTN2, CD34, fms, c-kit, HPK1, Bcl-2 family, p53, WAF1, RB, and WT) known to be involved in regulation of proliferation and differentiation of hemopoietic cells. (Abstract shortened by UMI.)M.Sc