Poster abstracts
Poster number 45 submitted by Woojin Han
Promoting a Novel Approach to Cellular Gene Expression Alteration (PANACEA)
Joseph Dong, Christopher Giromini, Woojin Han, Sonja Hatten, Ki Kim (University of Maryland), Autusa Pahlavan, Rajan Patel, Aniekanabasi Ufot, LeAnne Young (University of Maryland)
Abstract:
A novel method for delivering small interfering RNA (siRNA) to alter cellular gene expression was recently developed at the NIH. This method uses a modular vehicle consisting of a specific ligand coupled to a Hepatitis B Virus-derived RNA binding domain (HPV-RBD). The system enables researchers to deliver siRNAs to specific cell types through cell-specific receptor/ligand interactions. These interactions trigger cells to internalize the receptor/ligand complex via receptor-mediated endocytosis (RME). When thedelivery vehicle is internalized, so is the RNA cargo bound to HPV-RBD. The research objective is to develop and refine this novel small-molecule delivery system.Two novel recombinant delivery proteins are being developed: One with Interleukin-8 fused tothe HPV-RBD, the other with Machupo Virus GP1 joined to HPV-RBD. After incubating with specific siRNA cargo, the recombinant proteins will be exposed to CEM (a human T-cell line) or HeLa (epithelial) cell cultures. We predict the IL-8 vehicle will specifically deliver RNAs to T-cells through the IL8 receptors CXCR1 and CXCR2, while the Machupo virus GP1, which targets the ubiquitous transferrin receptor, will deliver RNAs to all cells. qRT-PCR will be used to measure changes in specific mRNA levels in boththe CEM and HeLa cells.A major limitation to safe, effective, and targeted delivery of therapeutic RNA to living cells is the harshness of conventional techniques. The gentle nature of this technology has the potential to overcome this limitation and could provide a platform for the expansion of personalized medicine
Keywords: siRNA, endocytosis, medicine