title: RNase P Ribozymes Inhibit the Replication of Human Cytomegalovirus by Targeting Essential Viral Capsid Proteins.
abstract: An engineered RNase P-based ribozyme variant, which was generated using the in vitro selection procedure, was used to target the overlapping mRNA region of two proteins essential for human cytomegalovirus (HCMV) replication: capsid assembly protein (AP) and protease (PR). In vitro studies showed that the generated variant, V718-A, cleaved the target AP mRNA sequence efficiently and its activity was about 60-fold higher than that of wild type ribozyme M1-A. Furthermore, we observed a reduction of 98%-99% in AP/PR expression and an inhibition of 50,000 fold in viral growth in cells with V718-A, while a 75% reduction in AP/PR expression and a 500-fold inhibition in viral growth was found in cells with M1-A. Examination of the antiviral effects of the generated ribozyme on the HCMV replication cycle suggested that viral DNA encapsidation was inhibited and as a consequence, viral capsid assembly was blocked when the expression of AP and PR was inhibited by the ribozyme. Thus, our study indicates that the generated ribozyme variant is highly effective in inhibiting HCMV gene expression and blocking viral replication, and suggests that engineered RNase P ribozyme can be potentially developed as a promising gene-targeting agent for anti-HCMV therapy.
authors.affiliation: Institute of Virology, School of Life Sciences, Nanjing University, Nanjing 210093, China. sin_angel@foxmail.com.|Taizhou Institute of Virology, Taizhou 225300, China. sin_angel@foxmail.com.|Jiangsu Affynigen Biotechnologies, Inc., Taizhou 225300, China. sin_angel@foxmail.com.
authors.forename: Zhu
authors.initials: Z
authors.lastname: Yang
authors.affiliation: School of Public Health, University of California, Berkeley, CA 94720, USA. mreeves@berkeley.edu.
authors.forename: Michael
authors.initials: M
authors.lastname: Reeves
authors.affiliation: The People's Hospital of Taizhou, Taizhou 225300, China. junye1965@hotmail.com.
authors.forename: Jun
authors.initials: J
authors.lastname: Ye
authors.affiliation: School of Public Health, University of California, Berkeley, CA 94720, USA. phong.trang@berkeley.edu.
authors.forename: Phong
authors.initials: P
authors.lastname: Trang
authors.affiliation: The People's Hospital of Taizhou, Taizhou 225300, China. lizhu1975@hotmail.com.
authors.forename: Li
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Program in Comparative Biochemistry, University of California, Berkeley, CA 94720, USA. jxsheng@berkeley.edu.
authors.forename: Jingxue
authors.initials: J
authors.lastname: Sheng
authors.affiliation: Taizhou Institute of Virology, Taizhou 225300, China. sin_angel@foxmail.com.|Jiangsu Affynigen Biotechnologies, Inc., Taizhou 225300, China. sin_angel@foxmail.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Institute of Virology, School of Life Sciences, Nanjing University, Nanjing 210093, China. kzen@nju.edu.cn.
authors.forename: Ke
authors.initials: K
authors.lastname: Zen
authors.affiliation: State Key Laboratory of Virology and College of Life Sciences, Wuhan University, Wuhan 430072, China. jwu@whu.edu.cn.
authors.forename: Jianguo
authors.initials: J
authors.lastname: Wu
authors.affiliation: School of Public Health, University of California, Berkeley, CA 94720, USA. liu_fy@berkeley.edu.|Program in Comparative Biochemistry, University of California, Berkeley, CA 94720, USA. liu_fy@berkeley.edu.
authors.forename: Fenyong
authors.initials: F
authors.lastname: Liu
mesh_terms: D036022:Capsid Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D003587:Cytomegalovirus / Q000235:genetics / Q000502:physiology*; D003586:Cytomegalovirus Infections / Q000235:genetics / Q000628:therapy / Q000821:virology*; D015967:Gene Expression Regulation, Viral; D006801:Humans; D016337:RNA, Catalytic / Q000235:genetics / Q000378:metabolism*; D043262:Ribonuclease P / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D036022:Capsid Proteins; D016337:RNA, Catalytic; D043262:Ribonuclease P
keywords: RNase P; cytomegalovirus; gene targeting; gene therapy; ribozyme