Functional cloning of genes that regulate hematopoiesis

Abstract

Abstract Retrovirus-mediated functional screening of cDNAs is feasible in identification of cDNAs in a variety of different ways. In this talk, I will present newly developed methods (SST-REX and FL-REX) as well as initial characterization of some cDNAs identified in the retrovirus-mediated expression screening. First, in functional screening of cDNA's involved in macrophage differentiation of M1 cells, we identified an anti-sense cDNA for MgcRacGAP which profoundly inhibited IL-6-driven differentiation of M1 cells. Interestingly, expression of MgcRacGAP by itself induced differentiation of HL60 cells into macrophages. Second, using a newly developed signal sequence trap method based on retrovirus-mediated expression (SST-REX), we identified a novel type I cytokine receptor δ1 which is similar to the common gamma subunit. Finally, I will also talk about another novel method FL-REX in which cDNAs are identified based on the subcellular localization of green fluorescent protein-fusion products.