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Inhibition of human cytomegalovirus major capsid protein expression and replication by ribonuclease P–associated external guide sequences

External guide sequences (EGSs) signify the short RNAs that induce ribonuclease P (RNase P), an enzyme responsible for processing the 5′ termini of tRNA, to specifically cleave a target mRNA by forming a precursor tRNA-like complex. Hence, the EGS technology may serve as a potential strategy for gen...

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Autores principales: Deng, Qiudi, Liu, Yujun, Li, Xin, Yan, Bin, Sun, Xu, Tang, Wei, Trang, Phong, Yang, Zhu, Gong, Hao, Wang, Yu, Lu, Jie, Chen, Jun, Xia, Chuan, Xing, Xiwen, Lu, Sangwei, Liu, Fenyong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6467005/
https://www.ncbi.nlm.nih.gov/pubmed/30803999
http://dx.doi.org/10.1261/rna.069682.118
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author Deng, Qiudi
Liu, Yujun
Li, Xin
Yan, Bin
Sun, Xu
Tang, Wei
Trang, Phong
Yang, Zhu
Gong, Hao
Wang, Yu
Lu, Jie
Chen, Jun
Xia, Chuan
Xing, Xiwen
Lu, Sangwei
Liu, Fenyong
author_facet Deng, Qiudi
Liu, Yujun
Li, Xin
Yan, Bin
Sun, Xu
Tang, Wei
Trang, Phong
Yang, Zhu
Gong, Hao
Wang, Yu
Lu, Jie
Chen, Jun
Xia, Chuan
Xing, Xiwen
Lu, Sangwei
Liu, Fenyong
author_sort Deng, Qiudi
collection PubMed
description External guide sequences (EGSs) signify the short RNAs that induce ribonuclease P (RNase P), an enzyme responsible for processing the 5′ termini of tRNA, to specifically cleave a target mRNA by forming a precursor tRNA-like complex. Hence, the EGS technology may serve as a potential strategy for gene-targeting therapy. Our previous studies have revealed that engineered EGS variants induced RNase P to efficiently hydrolyze target mRNAs. In the present research, an EGS variant was designed to be complementary to the mRNA coding for human cytomegalovirus (HCMV) major capsid protein (MCP), which is vital to form the viral capsid. In vitro, the EGS variant was about 80-fold more efficient in inducing human RNase P-mediated cleavage of the target mRNA than a natural tRNA-derived EGS. Moreover, the expressed variant and natural tRNA-originated EGSs led to a decrease of MCP expression by 98% and 73%–74% and a decrease of viral growth by about 10,000- and 200-fold in cells infected with HCMV, respectively. These results reveal direct evidence that the engineered EGS variant has higher efficiency in blocking the expression of HCMV genes and viral growth than the natural tRNA-originated EGS. Therefore, our findings imply that the EGS variant can be a potent candidate agent for the treatment of infections caused by HCMV.
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spelling pubmed-64670052020-05-01 Inhibition of human cytomegalovirus major capsid protein expression and replication by ribonuclease P–associated external guide sequences Deng, Qiudi Liu, Yujun Li, Xin Yan, Bin Sun, Xu Tang, Wei Trang, Phong Yang, Zhu Gong, Hao Wang, Yu Lu, Jie Chen, Jun Xia, Chuan Xing, Xiwen Lu, Sangwei Liu, Fenyong RNA Article External guide sequences (EGSs) signify the short RNAs that induce ribonuclease P (RNase P), an enzyme responsible for processing the 5′ termini of tRNA, to specifically cleave a target mRNA by forming a precursor tRNA-like complex. Hence, the EGS technology may serve as a potential strategy for gene-targeting therapy. Our previous studies have revealed that engineered EGS variants induced RNase P to efficiently hydrolyze target mRNAs. In the present research, an EGS variant was designed to be complementary to the mRNA coding for human cytomegalovirus (HCMV) major capsid protein (MCP), which is vital to form the viral capsid. In vitro, the EGS variant was about 80-fold more efficient in inducing human RNase P-mediated cleavage of the target mRNA than a natural tRNA-derived EGS. Moreover, the expressed variant and natural tRNA-originated EGSs led to a decrease of MCP expression by 98% and 73%–74% and a decrease of viral growth by about 10,000- and 200-fold in cells infected with HCMV, respectively. These results reveal direct evidence that the engineered EGS variant has higher efficiency in blocking the expression of HCMV genes and viral growth than the natural tRNA-originated EGS. Therefore, our findings imply that the EGS variant can be a potent candidate agent for the treatment of infections caused by HCMV. Cold Spring Harbor Laboratory Press 2019-05 /pmc/articles/PMC6467005/ /pubmed/30803999 http://dx.doi.org/10.1261/rna.069682.118 Text en © 2019 Deng et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.
spellingShingle Article
Deng, Qiudi
Liu, Yujun
Li, Xin
Yan, Bin
Sun, Xu
Tang, Wei
Trang, Phong
Yang, Zhu
Gong, Hao
Wang, Yu
Lu, Jie
Chen, Jun
Xia, Chuan
Xing, Xiwen
Lu, Sangwei
Liu, Fenyong
Inhibition of human cytomegalovirus major capsid protein expression and replication by ribonuclease P–associated external guide sequences
title Inhibition of human cytomegalovirus major capsid protein expression and replication by ribonuclease P–associated external guide sequences
title_full Inhibition of human cytomegalovirus major capsid protein expression and replication by ribonuclease P–associated external guide sequences
title_fullStr Inhibition of human cytomegalovirus major capsid protein expression and replication by ribonuclease P–associated external guide sequences
title_full_unstemmed Inhibition of human cytomegalovirus major capsid protein expression and replication by ribonuclease P–associated external guide sequences
title_short Inhibition of human cytomegalovirus major capsid protein expression and replication by ribonuclease P–associated external guide sequences
title_sort inhibition of human cytomegalovirus major capsid protein expression and replication by ribonuclease p–associated external guide sequences
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6467005/
https://www.ncbi.nlm.nih.gov/pubmed/30803999
http://dx.doi.org/10.1261/rna.069682.118
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