Abstract
The RNA cleavage activity of RNase P can be employed to decrease the levels of specific RNAs and to study their function or even to eradicate pathogens. Two different technologies have been developed to use RNase P as a tool for RNA knockdown. In one of these, an external guide sequence, which mimics a tRNA precursor, a well-known natural RNase P substrate, is used to target an RNA molecule for cleavage by endogenous RNase P. Alternatively, a guide sequence can be attached to M1 RNA, the (catalytic) RNase P RNA subunit of Escherichia coli. The guide sequence is specific for an RNA target, which is subsequently cleaved by the bacterial M1 RNA moiety. These approaches are applicable in both bacteria and eukaryotes. In this review, we will discuss the two technologies in which RNase P is used to reduce RNA expression levels.
Highlights
For many years, strategies to interfere with gene expression post-transcriptionally have been applied to study gene function in cultured cells
The application of these techniques will be illustrated by recent examples aimed at the down-regulation of microbial genes, genes involved in cancer and a host gene that is involved in HIV1 infection
In the case of F. tularensis and Y. pestis, the studies have been performed in E. coli and, before they can be used in therapeutic approaches, they need to be tested in the pathogenic strains
Summary
Strategies to interfere with gene expression post-transcriptionally have been applied to study gene function in cultured cells. 30 years ago, Altman and coworkers found that a model substrate, which resembles the acceptor stem of a pre-tRNA, was cleaved by E. coli RNase P in vitro [12]. RNase P is employed to knockdown specific RNAs. RNase P is employed to knockdown specific RNAs These techniques generally involve (external) guide sequences, which target specific RNAs and resemble the structure of the pre-tRNA substrates after base-pairing. We will discuss the current techniques for RNA down-regulation that are based on RNase P activity The application of these techniques will be illustrated by recent examples aimed at the down-regulation of microbial genes, genes involved in cancer and a (human) host gene that is involved in HIV1 infection
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