Medhat K Shier1, Hebatallah H Ali1, Mohammad S El-Wetidy1, Mohammad M Al-Qattan2, Mohamed B Al-Fageeh3. 1. College of Medicine Research Centre, King Saud University, Riyadh, Saudi Arabia. 2. Department of Surgery, College of Medicine, King Saud University, Riyadh, Saudi Arabia. 3. National Centre for Biotechnology, King Abdulaziz City for Science and Technology, Riyadh, Saudi Arabia.
Abstract
OBJECTIVE: To explore the effects of two different methodologies of RNA interference, namely small interfering RNA, and vector-based short hairpin RNA, on the expression levels of hepatitis C virus core RNA and protein of Saudi genotype 4 isolates. STUDY DESIGN: An experimental study. PLACE AND DURATION OF STUDY: Laboratories of the College of Medicine Research Center, King Saud University, Saudi Arabia, from January to December 2018. METHODOLOGY: Hepatitis C virus core small interfering RNA molecule and short hairpin RNA vector were designed against core region. Viral RNA expression was tested by RT-PCR; whereas, core protein was tested by flow cytometry and immunofluorescence. Results were statistically analysed by Chi-square analysis to calculate the p-value. RESULTS: Both molecules caused a reduction in core RNA and protein expression in infected cells. The effect of 100-pmol of small interfering RNA was more evident. For the vector-based short hairpin RNA, inhibition of core RNA expression was quite evident after 96 hours (p = 0.007). The results of flow cytometry and immunofluorescence showed a decline in core protein expression. The most dramatic effect was observed with 100-pmol small interfering RNA treatment of cells for 24 and 48 hours, which resulted in 63.5% and 91.1% core RNA expression reduction, respectively. CONCLUSION: RNA interference of hepatitis C virus core gene efficiently stopped viral replication and offer a promising therapeutic alternative against virus infection.
OBJECTIVE: To explore the effects of two different methodologies of RNA interference, namely small interfering RNA, and vector-based short hairpin RNA, on the expression levels of hepatitis C virus core RNA and protein of Saudi genotype 4 isolates. STUDY DESIGN: An experimental study. PLACE AND DURATION OF STUDY: Laboratories of the College of Medicine Research Center, King Saud University, Saudi Arabia, from January to December 2018. METHODOLOGY:Hepatitis C virus core small interfering RNA molecule and short hairpin RNA vector were designed against core region. Viral RNA expression was tested by RT-PCR; whereas, core protein was tested by flow cytometry and immunofluorescence. Results were statistically analysed by Chi-square analysis to calculate the p-value. RESULTS: Both molecules caused a reduction in core RNA and protein expression in infected cells. The effect of 100-pmol of small interfering RNA was more evident. For the vector-based short hairpin RNA, inhibition of core RNA expression was quite evident after 96 hours (p = 0.007). The results of flow cytometry and immunofluorescence showed a decline in core protein expression. The most dramatic effect was observed with 100-pmol small interfering RNA treatment of cells for 24 and 48 hours, which resulted in 63.5% and 91.1% core RNA expression reduction, respectively. CONCLUSION: RNA interference of hepatitis C virus core gene efficiently stopped viral replication and offer a promising therapeutic alternative against virus infection.
Authors: Reem M Aljowaie; Fahad N Almajhdi; Hebatallh H Ali; Mohammad S El-Wetidy; Medhat K Shier Journal: Cell Stress Chaperones Date: 2020-02-14 Impact factor: 3.667