Key result
Expression of a novel M1 RNA construct in mouse cells infected with HSV-1 reduced the levels of both TK mRNA and protein by approximately 80% compared to control cells.
A modified catalytic RNA subunit from E. coli RNase P can effectively and specifically inhibit viral gene expression in mammalian cells.
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Hypothesis-generating for RNase P ribozymes as antivirals; requires in vivo validation before any clinical consideration.
The catalytic RNA subunit (M1 RNA) of RNase P from Escherichia coli has been converted to an endoribonuclease that specifically cleaves the mRNA that encodes thymidine kinase (TK) of herpes simplex virus 1 (HSV-1). Covalent attachment to the 3' end of M1 RNA of a sequence complementary to TK mRNA results in very efficient cleavage of the target RNA in vitro. This reaction can be stimulated by proteins extracted from both E. coli and HeLa cells. When mouse cells in culture that express the novel RNA construct are infected with HSV-1, the levels of both TK mRNA and protein are reduced by approximately 80% as compared with cells that either do not express the novel RNA construct or express constructs with certain deletions that are known to abolish the catalytic activity of M1 RNA.
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Liu et al. (1995) studied HSV-1 infection (in vitro/cell culture). M1 RNA construct with complementary sequence to TK mRNA vs. Cells not expressing the construct or expressing constructs with deletions abolishing catalytic activity was evaluated on Levels of TK mRNA and protein. Expression of a novel M1 RNA construct in mouse cells infected with HSV-1 reduced the levels of both TK mRNA and protein by approximately 80% compared to control cells.
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