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Viruses. 2014 Jun 13;6(6):2376-91. doi: 10.3390/v6062376.

Engineered RNase P ribozymes effectively inhibit human cytomegalovirus gene expression and replication.

Author information

1
Institute of Virology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210093, China. nooney1986@163.com.
2
Program in Comparative Biochemistry, University of California, Berkeley, CA 94720, USA. giaphongvu@berkeley.edu.
3
Department of Gynecology, People's Hospital of Taizhou, Taizhou, Jiangsu 225300, China. huaqian1965@hotmail.com.
4
Program in Comparative Biochemistry, University of California, Berkeley, CA 94720, USA. yuchuan1022@berkeley.edu.
5
Taizhou Institute of Virology, Taizhou, Jiangsu 225300, China. sin_angel@foxmail.com.
6
School of Public Health, University of California, Berkeley, CA 94720, USA. mreeves@berkeley.edu.
7
Institute of Virology, School of Life Sciences, Nanjing University, Nanjing, Jiangsu 210093, China. kzen@nju.edu.cn.
8
Program in Comparative Biochemistry, University of California, Berkeley, CA 94720, USA. liu_fy@berkeley.edu.

Abstract

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.

PMID:
24932966
PMCID:
PMC4074932
DOI:
10.3390/v6062376
[Indexed for MEDLINE]
Free PMC Article

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