Journal of Biotechnology & Biomaterials
Open Access

Abstract

A classical technique for determining the function of a gene is to experimentally inhibit its expression in order to examine the resulting phenotype or effect on molecular endpoints and signaling pathways. RNA interference (RNAi) is one of the recent discoveries of a naturally occurring mechanism of gene regulation, initiated by the introduction of double stranded RNA into a cell. Synthetic short interfering RNAs (siRNAs) can be designed to silence the expression of specific genes bearing a particular target sequence and may potentially be presented as a therapeutic strategy for inhibiting transcriptional regulation of genes, which in such instances constitute a more attractive strategy than small molecule drugs due to its specificity. Low dose drug and siRNA combination studies are potential strategies for identifying synergistic targets, which facilitate reduction of undesired gene expression and/or cell growth depending on the research of interest. However, it is critical that any observed phenotypic change be confirmed at either the mRNA and/or protein level to determine the validity of the targeted genes. Quantitative real-time PCR (qPCR) is now extensively employed owing to its simplicity, wide dynamic range of quantification, sensitivity, and precision, for target-specific evaluation and validation of gene expression dynamics. We describe here a high-throughput screening of RNAi based gene silencing process and qPCR validation of specific transcript levels. In light of such advantageous applications, siRNA technology has become an ideal tool for studying gene function and holds the promise that siRNA-based therapeutic agents will soon be put to test in clinical trials.

Biography

Email: sukru.tuzmen@emu.edu.tr