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University of Wisconsin Projects

Project 1, “The De Novo Generation of Hematopoietic Stem Cells (Hscs) from Human ESC/iPCs and Somatic Cells.” Aims include:
Aim 1. Identify the hierarchy of mesodermal progenitors of vascular endothelial and hematopoietic lineages using human ES cell lines with targeted FOXF1, GATA-2, GATA-3, RUNX1, and SCL genes.
Aim 2. Determine the most critical molecular events leading to the formation of hematopoietic cells and HSCs during embryogenesis in mouse and human, and following in vitro differentiation of human pluripotent stem cells.
Aim 3. Develop technologies for de novo generation of HSCs from human ESC/iPSCs and somatic cells.
Project 2, “Cardiovascular Progenitors and Cardiomyocytes from Human Pluripotent Stem Cells and Somatic Cells.” Aims include:
Aim 1Isolate and define human embryonic cardiovascular progenitors derived from pluripotent stem cells.
Aim 2.  Reprogram human pluripotent stem cells and somatic cells using transcription factors to generate cardiovascular progenitors and ventricular myocytes.
Project 3, “Artificial Transcription Factors for Reprogramming/Transdifferentation.” Aims include:
Aim 1. Design ATFs that activate the expression of Oct4, Sox2 or Nanog in primary fibroblasts.
Aim 2. Perform genome-wide location analysis (ChIP-chip or ChIP-seq), CSI and transcriptome analysis to determine the gene targets and specificity of the designed ATFs.
Aim 3. Combine ATFs, alone or in combination with natural transcription factors, to induce pluripotency in primary fibroblasts.
Project 4, “Rapid In Vitro Generation of Affinity Reagents for Hematopoietic and Cardiovascular Precursors.” Aims include:
Aim 1. Develop methods for rapidly generating aptamers that bind to transmembrane protein targets with high affinity and specificity.
Aim 2. Develop methods to accelerate the discovery of aptamers with sub nanomolar affinities using high-throughput DNA sequencing technologies.
Aim 3.Generate high affinity aptamers for the surface markers of cardiovascular and hematopoietic precursors.
Genomics and Bioinformatics Core that will support each of the four proposed UO1 projects. Core aims include:
Aim 1. Isolate rare hematopoietic and cardiac precursor cell populations for genomic analysis.
Aim 2. Provide high throughput sequencing for RNA-Seq and ChiP-Seq.
Aim 3Provide Bioinformatics support for RNA-seq and ChIP-Seq.
Aim 4. Provide pools of heart and blood precursor cell cDNAs in lentiviral vectors.
Aim 5. Provide genetically modified human ES and iPS cells.


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