ABSTRACT
@#Gene manipulation tools have transformed biomedical research and improved the possibilities of their uses for therapeutic purposes. These tools have aided effective genomic modification in many organisms and have been successfully applied in biomedical engineering, biotechnology and biomedicine. They also shown a potential for therapeutic applications to alleviate genetic and non-genetic diseases. Small interfering RNA (siRNA) and clustered regularly inter-spaced short-palindromic repeat/associated-protein system (CRISPR/Cas) are two of the tools applied in genetic manipulation. This review aims to evaluate the molecular influence of siRNA and CRISPR/Cas as novel tools for genetic manipulations. This review discusses the molecular mechanism of siRNA and CRISPR/Cas, and the advantages and disadvantages of siRNA and CRISPR/Cas. This review also presents comparison between siRNA and CRISPR/Cas as potential tools for gene therapy. siRNA therapeutic applications occur through protein knockout without causing damage to cells. siRNA knocks down gene expression at the mRNA level, whereas CRISPR/Cas knocks out gene permanently at the DNA level. Inconclusion, gene manipulation tools have potential for applications that improve therapeutic strategies and plant-derived products, but ethical standards must be established before the clinical application of gene editing.
ABSTRACT
@#Introduction: Patients with Nasopharyngeal carcinoma (NPC) usually diagnosed at advanced cancer stage and recurrent case. Rac1 have become an emerging therapeutic target for metastasis cancer. This gene is critically involved in cell polarization and reactive oxygen species-mediated cell killing. This study aims to investigate the Rac1 activities in NPC/HK1 cell line using siRNA approach and evaluate the calcium deposition profile. Methods: The NPC/ HK1cells were transfected with Rac1-siRNA (siRac1) at concentrations of 50nM, 100nM and 200nM for 24 hours and stained with alizarin red s for calcium mineralization profile. Levels of Rac1 gene expression were measured via qRT-PCR followed by the time dependent assessment for 24, 48 and 72 hours. Results: Findings revealed that siRac1 concentrations of 200nM (p-value <0.02) and 100nM (p-value <0.016) had significant Rac1 suppression while 50nM (p-value <0.076) had the least suppression. On the other hand, from alizarin red S staining showed no significant changes for calcium mineralization activity on treated and control cells. However, siRac1 treated cells at 200nM showed presence of intracellular organelle swelling and loss of membrane integrity in 70% of the cells. This observation could possibly be linked to early sign of necrosis activity, hypoxia and disruption in intracellular calcium influx. Conclusion: This study suggest that Rac1 gene suppression might be involved in disruption of calcium deposition and reactive oxygen species-mediated NPC/HK1 cell killing. Further insight on the Rac1 molecular mechanism are needed to understand its potential role as therapeutic target for NPC