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Viruses 2014, 6(6), 2376-2391; doi:10.3390/v6062376

Engineered RNase P Ribozymes Effectively Inhibit Human Cytomegalovirus Gene Expression and Replication

Institute of Virology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210093, China
Taizhou Institute of Virology, Taizhou, Jiangsu 225300, China
Jiangsu Affynigen Biotechnologies, Inc., Taizhou, Jiangsu 225300, China
Program in Comparative Biochemistry, University of California, Berkeley, CA 94720, USA
Department of Gynecology, People's Hospital of Taizhou, Taizhou, Jiangsu 225300, China
School of Public Health, University of California, Berkeley, CA 94720, USA
These authors contribute equally to this study.
Author to whom correspondence should be addressed.
Received: 8 April 2014 / Revised: 19 May 2014 / Accepted: 23 May 2014 / Published: 13 June 2014
(This article belongs to the Special Issue Recent CMV Research)
View Full-Text   |   Download PDF [791 KB, uploaded 12 May 2015]   |  


RNase P ribozyme can be engineered to be a sequence-specific gene-targeting agent with promising application in both basic research and clinical settings. By using an in vitro selection system, we have previously generated RNase P ribozyme variants that have better catalytic activity in cleaving an mRNA sequence than the wild type ribozyme. In this study, one of the variants was used to target the mRNA encoding human cytomegalovirus (HCMV) essential transcription factor immediate-early protein 2 (IE2). The variant was able to cleave IE2 mRNA in vitro 50-fold better than the wild type ribozyme. A reduction of about 98% in IE2 expression and a reduction of 3500-fold in viral production was observed in HCMV-infected cells expressing the variant compared to a 75% reduction in IE2 expression and a 100-fold reduction in viral production in cells expressing the ribozyme derived from the wild type sequence. These results suggest that ribozyme variants that are selected to be highly active in vitro are also more effective in inhibiting the expression of their targets in cultured cells. Our study demonstrates that RNase P ribozyme variants are efficient in reducing HCMV gene expression and growth and are potentially useful for anti-viral therapeutic application. View Full-Text
Keywords: ribozyme; RNase P; gene targeting; cytomegalovirus ribozyme; RNase P; gene targeting; cytomegalovirus

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MDPI and ACS Style

Yang, Z.; Vu, G.-P.; Qian, H.; Chen, Y.-C.; Wang, Y.; Reeves, M.; Zen, K.; Liu, F. Engineered RNase P Ribozymes Effectively Inhibit Human Cytomegalovirus Gene Expression and Replication. Viruses 2014, 6, 2376-2391.

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