Effect of RNA Interference on the Hepatitis C Virus Core Expression in HepG2/C3A Cells Using Genotype 4 Isolates from Saudi Patients
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.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2019 |
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| Erschienen: |
2019 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:29 |
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| Enthalten in: |
Journal of the College of Physicians and Surgeons--Pakistan : JCPSP - 29(2019), 9 vom: 01. Sept., Seite 852-859 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Shier, Medhat K [VerfasserIn] |
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| Links: |
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| Themen: |
Journal Article |
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| Anmerkungen: |
Date Completed 06.02.2020 Date Revised 06.02.2020 published: Print Citation Status MEDLINE |
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| doi: |
10.29271/jcpsp.2019.09.852 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM300650396 |
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| 100 | 1 | |a Shier, Medhat K |e verfasserin |4 aut | |
| 245 | 1 | 0 | |a Effect of RNA Interference on the Hepatitis C Virus Core Expression in HepG2/C3A Cells Using Genotype 4 Isolates from Saudi Patients |
| 264 | 1 | |c 2019 | |
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| 500 | |a Date Completed 06.02.2020 | ||
| 500 | |a Date Revised 06.02.2020 | ||
| 500 | |a published: Print | ||
| 500 | |a Citation Status MEDLINE | ||
| 520 | |a 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 | ||
| 520 | |a STUDY DESIGN: An experimental study | ||
| 520 | |a PLACE AND DURATION OF STUDY: Laboratories of the College of Medicine Research Center, King Saud University, Saudi Arabia, from January to December 2018 | ||
| 520 | |a 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 | ||
| 520 | |a 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 | ||
| 520 | |a CONCLUSION: RNA interference of hepatitis C virus core gene efficiently stopped viral replication and offer a promising therapeutic alternative against virus infection | ||
| 650 | 4 | |a Journal Article | |
| 650 | 7 | |a RNA, Small Interfering |2 NLM | |
| 650 | 7 | |a RNA, Viral |2 NLM | |
| 650 | 7 | |a Viral Core Proteins |2 NLM | |
| 700 | 1 | |a Ali, Hebatallah H |e verfasserin |4 aut | |
| 700 | 1 | |a El-Wetidy, Mohammad S |e verfasserin |4 aut | |
| 700 | 1 | |a Al-Qattan, Mohammad M |e verfasserin |4 aut | |
| 700 | 1 | |a Al-Fageeh, Mohamed B |e verfasserin |4 aut | |
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