This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Dr. M.R.S. Krishnamani, a Post-Doctoral fellow sponsored by RCMI worked on the project titled Innovative Therapeutic Approaches for Sickle Cell Disease . During this time, Dr. Krishnamani was trained in molecular hematology research, and he focused his research and development in this area. Below we describe Dr. Krishnamani s research work. Sickle Cell Disease (SCD) is an inherited chronic hemolytic anemia due to a substitution of glutamic acid with valine at the position of amino acid six in the ? globin gene chain. Currently, there is no cure for SCD with the exception of bone marrow transplantation. There is a basic need to develop innovative therapeutic approaches to cure and reduce the risks of complications resulting from SCD. Gene therapy is the best candidate for a cure, with the development of an ideal vector for suitable globin gene delivery. In this study, we are creating plasmid based vectors with necessary and essential elements for expression and propagation inside k562 cell lines. One of the important elements is the introduction of a human Autonomous Replicating Sequence (ARS) which putatively assist in the successful replication of the plasmid inside mammalian cells. We were successful in (1) creating several plasmid constructs for ?-globin gene delivery with an aim to enrich for fetal hemoglobin (which has antisickling properties) inside k562 cells. These vectors were transfected into K562 cell lines and transfection efficiency and fetal hemoglobin levels were measured in the transfected cells.
SPECIFIC AIMS The overall aim of the Post-Doctoral fellowship is to assist the Post-Doctoral Fellow to achieve: 1) Research Focus in the Hemoglobinopathies 2) Gain technological experience in molecular hematology to produce gene therapy vectors to cure sickle cell disease.
Showing the most recent 10 out of 225 publications
