An Engineered RNase P Ribozyme Effectively Reduces Human Coronavirus 229E Gene Expression and Growth in Human Cells

The human coronavirus 229E (HCoV-229E) is a member of the human coronavirus family that includes SARS-CoV-2, the causative agent of COVID-19. Developing antiviral strategies and compounds is crucial to treat and prevent HCoV-229E infections and the associated diseases. Ribozymes derived from ribonuc...

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Main Authors: Yujun Liu, Bin Yan, Hao Gong, Fenyong Liu
Format: Article
Language:English
Published: MDPI AG 2025-05-01
Series:Zoonotic Diseases
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Online Access:https://www.mdpi.com/2813-0227/5/2/12
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author Yujun Liu
Bin Yan
Hao Gong
Fenyong Liu
author_facet Yujun Liu
Bin Yan
Hao Gong
Fenyong Liu
author_sort Yujun Liu
collection DOAJ
description The human coronavirus 229E (HCoV-229E) is a member of the human coronavirus family that includes SARS-CoV-2, the causative agent of COVID-19. Developing antiviral strategies and compounds is crucial to treat and prevent HCoV-229E infections and the associated diseases. Ribozymes derived from ribonuclease P (RNase P) catalytic RNA represent a novel class of promising gene-targeting agents by cleaving their target mRNA and knocking down the expression of the target mRNA. However, it has not been reported whether RNase P ribozymes block the infection and replication of HCoV-229E. We report here the engineering of an anti-HCoV-229E RNase P ribozyme to target an overlapping region of viral genomic RNA and the mRNA encoding the nucleocapsid (N) protein, which is vital for viral replication and growth. The engineered ribozyme actively hydrolyzed the viral RNA target in vitro. HCoV-229E-infected cells expressing the engineered, catalytically active ribozyme exhibited a reduction of about 85% in viral RNA levels and N protein expression, and a reduction of about 750-fold in infectious particle production, compared to cells expressing no ribozymes or a control, catalytically inactive ribozyme. Our study provides the first direct evidence of the therapeutic potential of RNase P ribozymes against human coronaviruses such as HCoV-229E.
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spelling doaj-art-9ba6b3bbc827461f86cabdeeba291f582025-08-20T03:26:57ZengMDPI AGZoonotic Diseases2813-02272025-05-01521210.3390/zoonoticdis5020012An Engineered RNase P Ribozyme Effectively Reduces Human Coronavirus 229E Gene Expression and Growth in Human CellsYujun Liu0Bin Yan1Hao Gong2Fenyong Liu3School of Public Health, University of California, Berkeley, CA 94720, USASchool of Public Health, University of California, Berkeley, CA 94720, USASchool of Public Health, University of California, Berkeley, CA 94720, USASchool of Public Health, University of California, Berkeley, CA 94720, USAThe human coronavirus 229E (HCoV-229E) is a member of the human coronavirus family that includes SARS-CoV-2, the causative agent of COVID-19. Developing antiviral strategies and compounds is crucial to treat and prevent HCoV-229E infections and the associated diseases. Ribozymes derived from ribonuclease P (RNase P) catalytic RNA represent a novel class of promising gene-targeting agents by cleaving their target mRNA and knocking down the expression of the target mRNA. However, it has not been reported whether RNase P ribozymes block the infection and replication of HCoV-229E. We report here the engineering of an anti-HCoV-229E RNase P ribozyme to target an overlapping region of viral genomic RNA and the mRNA encoding the nucleocapsid (N) protein, which is vital for viral replication and growth. The engineered ribozyme actively hydrolyzed the viral RNA target in vitro. HCoV-229E-infected cells expressing the engineered, catalytically active ribozyme exhibited a reduction of about 85% in viral RNA levels and N protein expression, and a reduction of about 750-fold in infectious particle production, compared to cells expressing no ribozymes or a control, catalytically inactive ribozyme. Our study provides the first direct evidence of the therapeutic potential of RNase P ribozymes against human coronaviruses such as HCoV-229E.https://www.mdpi.com/2813-0227/5/2/12coronavirushuman coronavirus 229ESARS-CoV-2ribozymeRNase Pgene targeting
spellingShingle Yujun Liu
Bin Yan
Hao Gong
Fenyong Liu
An Engineered RNase P Ribozyme Effectively Reduces Human Coronavirus 229E Gene Expression and Growth in Human Cells
Zoonotic Diseases
coronavirus
human coronavirus 229E
SARS-CoV-2
ribozyme
RNase P
gene targeting
title An Engineered RNase P Ribozyme Effectively Reduces Human Coronavirus 229E Gene Expression and Growth in Human Cells
title_full An Engineered RNase P Ribozyme Effectively Reduces Human Coronavirus 229E Gene Expression and Growth in Human Cells
title_fullStr An Engineered RNase P Ribozyme Effectively Reduces Human Coronavirus 229E Gene Expression and Growth in Human Cells
title_full_unstemmed An Engineered RNase P Ribozyme Effectively Reduces Human Coronavirus 229E Gene Expression and Growth in Human Cells
title_short An Engineered RNase P Ribozyme Effectively Reduces Human Coronavirus 229E Gene Expression and Growth in Human Cells
title_sort engineered rnase p ribozyme effectively reduces human coronavirus 229e gene expression and growth in human cells
topic coronavirus
human coronavirus 229E
SARS-CoV-2
ribozyme
RNase P
gene targeting
url https://www.mdpi.com/2813-0227/5/2/12
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