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Effective Inhibition of Human Cytomegalovirus Gene Expression and Replication by a Ribozyme Derived from the Catalytic RNA Subunit of RNase P from Escherichia coli
Phong Trang, Manfred Lee, Edward Nepomuceno, Joe Kim, Hua Zhu and Fenyong Liu
Proceedings of the National Academy of Sciences of the United States of America
Vol. 97, No. 11 (May 23, 2000), pp. 5812-5817
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/122539
Page Count: 6
You can always find the topics here!Topics: RNA, Messenger RNA, Cell lines, Infections, Cell growth, Gels, Cytomegalovirus, Actins, Antivirals, Viruses
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A sequence-specific ribozyme (M1GS RNA) derived from the catalytic RNA subunit of RNase P from Escherichia coli was used to target the overlapping exon 3 region of the mRNAs encoding the major transcription regulatory proteins IE1 and IE2 of human cytomegalovirus. A reduction of more than 80% in the expression levels of IE1 and IE2 and a reduction of about 150-fold in viral growth were observed in human cells that stably expressed the ribozyme. In contrast, a reduction of less than 10% in the IE1/IE2 expression and viral growth was observed in cells that either did not express the ribozyme or produced a "disabled" ribozyme that carried mutations that abolished its catalytic activity. Examination of the expression of several other viral early and late genes in the cells that expressed the M1GS ribozyme further revealed an overall reduction of at least 80% in their expression. These results are consistent with the notion that the antiviral effects in these cells are due to the fact that the ribozyme specifically inhibits the expression of IE1 and IE2 and, consequently, abolishes the expression of viral early and late genes as well as viral growth. Our study is the first, to our knowledge, to use M1GS ribozyme for inhibiting human cytomegalovirus replication and demonstrates the utility of this ribozyme for antiviral applications.
Proceedings of the National Academy of Sciences of the United States of America © 2000 National Academy of Sciences