Project description:To dissect the metabolic state of fetal liver HSCs, we have developed a genetically encoded fluorescent sensor (SoNar) for tracking cytosolic NADH and NAD+ redox states and constructed the transgenic mouse. Based on the NADH/NAD+ ratio, we have sorted SoNar-high and SoNar-low fetal liver blood cells to employ whole genome microarray expression profiling as a discovery platform to identify genes with the potential to regulate fetal liver HSC metabolism.
Project description:To dissect the metabolic state of B-ALL cells, we have developed a genetically encoded fluorescent sensor (SoNar) for tracking cytosolic NADH and NAD+ redox states and constructed the transgenic mouse. Based on the NADH/NAD+ ratio, we have sorted SoNar-high and SoNar-low B-ALL cells to employ whole genome microarray expression profiling as a discovery platform to identify genes with the potential to regulate the metabolism of B-ALL cells.
Project description:To dissect the metabolic state of B-ALL cells, we have developed a genetically encoded fluorescent sensor (SoNar) for tracking cytosolic NADH and NAD+ redox states and constructed the transgenic mouse. Based on the NADH/NAD+ ratio, we have sorted SoNar-high and SoNar-low B-ALL cells to employ whole genome microarray expression profiling as a discovery platform to identify genes with the potential to regulate the metabolism of B-ALL cells.
Project description:To investigate the impact of maternal nutrient restriction on gene expression in the fetal liver, we profiled gene expression in the liver of Japanese Black fetal calves in high and low nutrient condition of pregnant maternal cows .
Project description:RNA metabolism is controlled by an expanding yet incomplete catalog of RNA binding proteins (RBPs), many of which lack characterized RNA binding domains. Approaches to expand the RBP repertoire to discover non-canonical RBPs are currently needed. Here, HaloTag fusion pull-down of twelve nuclear and cytoplasmic RBPs followed by quantitative mass-spectrometry (MS) demonstrates that proteins interacting with multiple RBPs in an RNA-dependent manner are enriched for RBPs. This motivated SONAR, a computational approach that predicts RNA binding activity by analyzing large-scale affinity precipitation-MS protein-protein interactomes. Without relying on sequence or structure information, SONAR identifies 1923 human, 489 fly and 745 yeast RBPs, including over 100 human candidate RBPs that contain zinc finger domains. Enhanced CLIP confirms RNA binding activity and identifies transcriptome-wide RNA binding sites for SONAR-predicted RBPs, revealing unexpected RNA binding activity for disease-relevant proteins and DNA binding proteins.