The development of single cell microfluidic technologies for analysis of microRNA expression and epigenetic state with application to hematopoietic stem cells

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This project will create new and robust technology to assess the transcriptional heterogeneity of single cells at a genomic level. The prototype example to be used as a test case is the stem cell of the hematopoietic system where compelling evidence now indicates that different fate decisions are determined in part by mechanisms that are self-propagating through many self-renewal divisions. Understanding the molecular basis of these mechanisms is fundamental to current intense efforts to expand hematopoietic stem cells ex vivo, to develop safer and more effective gene therapy protocols, and to elucidate mechanisms of leukemogenesis in individual patients. The technology to be developed, validated and tested in proof-of-principle experiments on highly purified populations of normal mouse and human hematopoietic stem cells will enable microRNA (miRNA) expression and epigenomic marks to be examined on hundreds of individual cells from small samples with high fidelity and at low cost. MiRNA analyses will utilize on-chip extraction, barcoding and PCR amplification