ArticleNucleic acids research2002
A functional gene discovery in the Fas-mediated pathway to apoptosis by analysis of transiently expressed randomized hybrid-ribozyme libraries.
Article in Nucleic acids research, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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13 citing papers in PubMed, 38 citations in OpenAlex.
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- A structural analysis of in vitro catalytic activities of hammerhead ribozymes.BMC bioinformatics · 2007Article
- Complete, gene-specific siRNA libraries: production and expression in mammalian cells.RNA (New York, N.Y.) · 2005Article
- Induction and characterization of taxol-resistance phenotypes with a transiently expressed artificial transcriptional activator library.Nucleic acids research · 2004Article
- Expressing functional siRNAs in mammalian cells using convergent transcription.BMC biotechnology · 2003Article
- Short hairpin type of dsRNAs that are controlled by tRNA(Val) promoter significantly induce RNAi-mediated gene silencing in the cytoplasm of human cells.Nucleic acids research · 2003Article
- Confirmation by FRET in individual living cells of the absence of significant amyloid beta -mediated caspase 8 activation.Proceedings of the National Academy of Sciences of the United States of America · 2002Article
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2 authors at 1 institution in 1 country.
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Abstract
The sequence of much of the human genome is now available and the next goal is to identify functional genes and to clarify their roles. We have recently developed a novel system for isolation of genes in the Fas- and TNF-alpha-mediated pathways to apoptosis using poly(A)-connected hammerhead ribozyme libraries with randomized substrate-binding arms at both the 5' and 3' ends of ribozymes. The transcripts of these hybrid ribozymes have a poly(A) motif that can recruit RNA helicases and, thus, they can effectively attack target sites. In the previous studies, hybrid ribozymes were stably expressed. In order to save selection times, in this study we adopted transiently expressed hybrid ribozymes. In the case of Fas-mediated apoptosis, when we transiently introduced these hybrid-ribozyme libraries into Fas-expressing HeLa cells, we were able to isolate surviving clones that were resistant to or exhibited a delay in Fas-mediated apoptosis. We identified many pro-apoptotic genes and novel genes using this strategy with these transiently expressed hybrid-ribozyme libraries. In contrast, we identified significantly smaller numbers of candidate genes using conventional ribozyme libraries that were expressed transiently. Thus, when changes of a particular phenotype occur within a short period of time, our gene discovery system based on transiently expressed hybrid-ribozyme libraries should also be useful for the rapid identification of functional genes in the post-genome era.
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