ArticleMolecular and cellular biology1995
Selection of efficient cleavage sites in target RNAs by using a ribozyme expression library.
Article in Molecular and cellular biology, 1995. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 34 papers.
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Who cites it
34 citing papers in PubMed, 148 citations in OpenAlex.
- Enhanced hammerhead ribozyme turnover rates: Reevaluating therapeutic space for small catalytic RNAs.Molecular therapy. Nucleic acids · 2025Article
- Systematic Screening, Rational Development, and Initial Optimization of Efficacious RNA Silencing Agents for Human Rod Opsin Therapeutics.Translational vision science & technology · 2019Article
- A cellular high-throughput screening approach for therapeutic trans-cleaving ribozymes and RNAi against arbitrary mRNA disease targets.Experimental eye research · 2016Article
- Relieving bottlenecks in RNA drug discovery for retinal diseases.Advances in experimental medicine and biology · 2012Review
- Rapid, cell-based toxicity screen of potentially therapeutic post-transcriptional gene silencing agents.Experimental eye research · 2011Article
- Variables and strategies in development of therapeutic post-transcriptional gene silencing agents.Journal of ophthalmology · 2011Article
- Effective suppression of Dengue fever virus in mosquito cell cultures using retroviral transduction of hammerhead ribozymes targeting the viral genome.Virology journal · 2009Article
- Development of lead hammerhead ribozyme candidates against human rod opsin mRNA for retinal degeneration therapy.Experimental eye research · 2009Article
- Bottlenecks in development of retinal therapeutic post-transcriptional gene silencing agents.Vision research · 2008Review
- Identification of cellular cofactors for human immunodeficiency virus replication via a ribozyme-based genomics approach.Journal of virology · 2004Article
- Oligonucleotide-directed site-specific integration of high complexity libraries into ssDNA templates.Nucleic acids research · 2004Article
- Specific hammerhead ribozymes reduce synthesis of cation-independent mannose 6-phosphate receptor mRNA and protein.Gene therapy · 2003Article
- E6AP gene suppression and characterization with in vitro selected hammerhead ribozymes.Cancer gene therapy · 2003Article
- Selection of targets and the most efficient hairpin ribozymes for inactivation of mRNAs using a self-cleaving RNA library.EMBO reports · 2001Article
- Identifying ribozyme-accessible sites using NUH triplet-targeting gapmers.Nucleic acids research · 2001Article
- A selection system for identifying accessible sites in target RNAs.RNA (New York, N.Y.) · 2001Article
- Mapping of RNA accessible sites by extension of random oligonucleotide libraries with reverse transcriptase.RNA (New York, N.Y.) · 2001Article
- Secondary structure prediction and in vitro accessibility of mRNA as tools in the selection of target sites for ribozymes.Nucleic acids research · 2000Article
- RNA accessibility prediction: a theoretical approach is consistent with experimental studies in cell extracts.Nucleic acids research · 2000Article
- Ribozyme-mediated resistance to rice dwarf virus and the transgene silencing in the progeny of transgenic rice plants.Transgenic research · 2000Article
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Authors and funding
2 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Inactivation of gene expression by antisense mechanisms in general and by ribozymes in particular is a powerful technique for studying the function of a gene product. We have designed a strategy for expression of ribozymes, for selection of accessible cleavage sites in target RNAs, and for isolation of ribozymes from a library of random sequences flanking the unique sequence of a hammerhead. The expression cassette for ribozyme genes is based on adenovirus-associated RNA. Alternatively, we used polymerase III or the T7 phage transcription machinery. The ribozyme sequences are positioned in the center of a stable stem-loop structure, allowing for a correctly folded ribozyme region within the expressed RNA. A library of ribozyme genes with random sequences of 13 nucleotides on both sides of the hammerhead was generated. As an example, ribozymes which are specific for seven sites within the mRNA or nuclear RNA of human growth hormone were selected and identified. Sequencing of ribozyme genes reamplified from the library confirmed not only the predicted cleavage sites but also the presence of different ribozyme variants in the library. In a test of the ribozyme variants for repression of growth hormone synthesis in a cellular assay, the strongest effect (more than 99% inhibition) was found for the variant with the shortest stretch of complementarity (7 and 8 nucleotides on either side) to the target RNA. This basic strategy seems to be applicable to the selection of suitable target sites and to the isolation of corresponding ribozymes for any mRNA of interest.
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