Cell-penetrating peptides as drug delivery system for SiRNA.

• Main Author: Meriska Sudarmadji.
• Other Authors: Bjoern Holger Neu
• Format: Final Year Project
• Language: English
• Published: 2012
• Subjects: DRNTU::Engineering::Chemical engineering::Biotechnology
• Online Access: http://hdl.handle.net/10356/50007
• Institution: Nanyang Technological University

In recent years, siRNA has gained a lot of attention in biomedical research field due to its ability to modulate specific protein expressions – a crucial factor in treating inflammation-related diseases. In the cytoplasm, siRNA assembles into RNA-induced silencing complex (RISC), facilitating the removal of target mRNA molecules that bind to RISC antisense strand. Therefore, siRNA is very potent in silencing any genes of interest. Nevertheless, siRNA’s potency is hampered by its inability to penetrate cell membrane. Thus, finding a suitable delivery vehicle to the cell is pivotal for siRNA to exert its effect. In this study, human calcitonin-derived peptides (hCT) and Endo-Porter – two novel cell-penetrating peptides (CPPs) – would be assessed as siRNA delivery vehicles. Their concentration ratios with siRNA and incubation time would be optimized in order to have an effective siRNA uptake. Thus, confocal laser scanning microscope (CLSM) and flow cytometry are employed to investigate the optimum parameters. Furthermore, to foresee hCT and Endo-Porter’s possible applications in clinical settings, their cytotoxicity toward the cells is studied through MTT assay and Trypan Blue exclusion method. Having demonstrated a more effective uptake of siGLO – a model siRNA – than hCT, along with a relatively low cytotoxicity, Endo-Porter is prospective for further investigation as a delivery vehicle for siRNA to treat inflammation-related diseases.