SUMMER RESEARCH TRAINING IN MEMBRANE SCIENCE AND TECHNOLOGY 2008 NSF REU SITE PROGRAM at the UNIVERSITY OF CINCINNATI
The Department of Pharmacology & Cell Biophysics, College of Medicine is pleased to offer this research project as part of the 2008 summer NSF-REU Site Program administered by the Department of Pharmacology & Cell Biophysics.  Students interested in this project are urged to contact Professor Jones to discover more about the project, learn what your responsibilities will be during the ten-week research training program.
Project #:  08-011 Faculty Supervisor/Mentor: W. Keith Jones. Ph.D., Associate Professor, Pharmacology & Cell Biophysics College of Medicine Email:  joneswk@uc.edu
Polymer-Facilitated Delivery of Nucleic Acids Across Bio-Membranes for Modulation of Gene Expression
General background and significance of the project: Ischemia/reperfusion (I/R) injury in biological tissues is a complex phenomenon that includes production of reactive oxygen species, nitric oxide calcium overload, cytokine release and signaling, expression/secretion of chemokines and chemoattractant molecules, the resulting infiltration and inflammation and extracellular matrix remodeling. NF-kappaB is a transcription factor that is implicated in the regulation of many genes that play pivotal roles in multiple aspects of cellular and tissue inflammation and I/R injury. We have shown that blocking NF-kappaB activation reduces death of heart muscle cells by 65%, an effect that is dependent upon dysregulation of NF-kappaB-dependent genes. In collaboration with Dr. Theresa Reineke (Dept. Chemistry) we have developed novel decoy DNA molecules that block NF-kappaB activation and are developing siRNA molecules to block individual NF-kappaB subunits. We are testing the ability of these novel non-viral polymers to deliver decoys and siRNA/shRNA across bio-membranes of heart muscle cells as a model system. The goal is to employ these to experimentally perturb signaling and gene expression in these cells to dissect the transcriptional signaling network.
Brief description of proposed research and activities for the 10-week REU period: The REU student will learn recombinant DNA techniques necessary for working with DNA constructs that contain transcription factor decoys and transgenes that express siRNAs (shRNAs). The REU student will prepare several varieties of DNA or siRNA/polymer complexes and implement experiments to deliver these reagents to isolated cardiomyocytes and/or the heart in vivo to determine the efficiency and efficacy of delivery across this model bio-membrane.
What the REU Student can gain from participating in this project: The REU student will learn basic molecular biology techniques and will become skilled with cell culture and dose response studies using cultured cells. Experimental design and scientific method will be emphasized. The REU student will be challenged and guided to read the biomedical scientific literature, will be exposed to successful scientists at various levels in the PIs laboratory (graduate students, postdoctoral fellows, junior faculty and faculty). The REU student will be expected to participate in journal clubs and interact on a daily basis with these scientists. The REU student will be guided in scientific writing and will have the opportunity to earn co-authorship when the studies are published.