This is a SERCA application for Dr. Susan Farmer to continue her training as a basic research and comparative medicine scientist. Support for this project would be provided by my primary sponsor, Dr. Tim Townes, Professor of Biochemistry and Molecular Genetics, and my secondary sponsor, Dr. Russell Lindsey, Chairman of the Department of Comparative Medicine. I would have access to the excellent resources and environment of both of these departments and the Animal Resources Program at the University of Alabama at Birmingham. As a Fellow in the Department of Comparative Medicine at UAB, I have completed training in clinical laboratory animal medicine, participated in a variety of research projects, and been admitted to candidacy for the PhD degree in Molecular and Cellular Pathology. My primary interests are mammalian embryonic development, molecular genetics, the pathogenesis of genetic disease, and animal models for human disease. My long term goal is to become a successful research scientist and teacher. The goal of this research project is to characterize the structure and function of the gene for a transcription factor, locus control region factor 1 (LCR-F1). Its cDNA was recently isolated based on the ability of LCR-F1 to bind duplicated AP1-like sites found in the promoters of globin genes and in the (3-globin locus control region (LCR). Proteins which bind these sites may function in transcriptional regulation of the a-like globin genes. Since LCR-F1 is a CNC domain family member, it may also be important in development. We have isolated the mouse genomic clone and used it to construct a gene targeting vector for producing mouse embryonic stem cells with deletions of LCR-F1 coding sequences. Chimeric mice have been made from these cells and are these animals are being bred to produce homozygous mutants to assess the effect of loss of LCR-F1 function. Loss of this protein in all cells could be lethal; therefore, to define the effects of LCR-F1 loss to erythroid cells, we plan to make mice with an erythroid-specific knockout of the gene using the Cre-lox system. We will also study regulation of expression of LCR-F1 by characterizing its promoter region. Finally, we will isolate other CNC family members for similar future studies. This work should provide insight into the role of LCR-F1 both in development, hematopoiesis, and globin gene expression. This-work should also provide new information on the CNC domain family of proteins. This SERCA would combine outstanding training in laboratory animal medicine, comparative medicine, and molecular genetics. The research project will provide both the training necessary for me to become an independent investigator and new information on hematopoiesis, transcriptional regulation, and development.
