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Development of a Hepatitis B Virus Reporter System to Monitor the Early Stages of the Replication Cycle
Published on: February 1, 2017
Effect of RNA Interference on the Hepatitis C Virus Core Expression in HepG2/C3A Cells Using Genotype 4 Isolates from
Medhat K Shier1, Hebatallah H Ali1, Mohammad S El-Wetidy1
1College of Medicine Research Centre, King Saud University, Riyadh, Saudi Arabia.
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.
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