David Ginsburg is interested in understanding the components of the blood-clotting system and how disturbances in their function lead to human bleeding and blood-clotting disorders.
Using TALEN Technology to Modify COPII Genes in Human Cell Lines
About 30% of the human genome encodes proteins that rely on the proper function of the secretory pathway. ER-to-Golgi transport represents the entry point of the secretory pathway and is mediated by the evolutionarily conserved coatamer complex II (COPII). Mutations in several COPII genes lead to distinct genetic diseases in human patients, with often markedly different findings in genetically engineered mice deficient in the orthologous protein. This project will aim to use a new genome-editing technology using TALENs to specifically tailor individual COPII genes in relevant cell types of human origin.
Analysis of ENU-Induced Mutations in Mice Affecting Blood Clotting
Our lab is focused on studying the genetics of blood coagulation. One particular blood-clotting disease, called venous thromboemboembolism, affects ~300,000 individuals/year in the United States. A polymorphism in the coagulation factor V gene, called Factor V Leiden (FVL), is the most common genetic risk factor for venous thrombosis, although it displays only 10% penetrance. We previously demonstrated synthetic lethality between homozygosity for FVL (FVQ/Q) and heterozygous tissue factor pathway inhibitor deficiency (TFPI+/–) in mice. To identify novel thrombosis modifier genes, we recently performed a mouse whole-genome ENU mutagenesis screen. By screening through 7,128 mice, we identified 98 putative thrombosis modifiers. We validated 14 of these and created mouse lines harboring these mutations. To expedite the identification of the suppressing mutations, we have Next-Gen whole-exome sequenced DNA from a mouse of each of the lines. We are presently screening through candidate mutations identified as a result of the Next-Gen sequencing. The identification of these suppressor mutations should provide novel insights into hemostatic regulation and will provide a starting point for future experiments aimed at determining the precise role of these genes in venous thrombosis and other hemostatic disorders. During this project, the student will participate in screening and validating the candidate modifier genes using mouse models.