Poster abstracts
Poster number 33 submitted by Joseph Dong
Promoting A Novel Approach to Cellular Gene Expression Alteration
Joseph Dong, Christopher Giromini, Woojin Han, Leanne Young (UMD), Aniekanabasi Ufot, Ki Kim, Rajan Patel, Autusa Pahlavan, Sonya Hatten (UMD), Johnathan Dinman (UMD)
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 the delivery 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 to the 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 both the 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: gene expression, RNA interference, receptor mediated endocytosis