ArticleNucleic acids research2017
The influence of the 5΄-terminal nucleotide on AgoshRNA activity and biogenesis: importance of the polymerase III transcription initiation site.
Article in Nucleic acids research, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 16 papers.
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Who cites it
16 citing papers in PubMed, 21 citations in OpenAlex.
- Design of antiviral AGO2-dependent short hairpin RNAs.Virologica Sinica · 2024Article
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- Combining protein and RNA quantification to evaluate promoter activity by using dual-color fluorescent reporting systems.Bioscience reports · 2021Article
- CRISPR therapy towards an HIV cure.Briefings in functional genomics · 2020Article
- Gene knockdown in malaria parasites via non-canonical RNAi.Nucleic acids research · 2020Article
- Short Hairpin RNAs for Strand-Specific Small Interfering RNA Production.Frontiers in bioengineering and biotechnology · 2020Article
- Article
- Influence of a 3' Terminal Ribozyme on AgoshRNA Biogenesis and Activity.Molecular therapy. Nucleic acids · 2019Article
- Efficient Knockdown and Lack of Passenger Strand Activity by Dicer-Independent shRNAs Expressed from Pol II-Driven MicroRNA Scaffolds.Molecular therapy. Nucleic acids · 2019Article
- RNA Polymerase II Activity of Type 3 Pol III Promoters.Molecular therapy. Nucleic acids · 2018Article
- Delineation of the Exact Transcription Termination Signal for Type 3 Polymerase III.Molecular therapy. Nucleic acids · 2018Article
- Influence of the loop size and nucleotide composition on AgoshRNA biogenesis and activity.RNA biology · 2017Article
- Dicer-independent processing of small RNA duplexes: mechanistic insights and applications.Nucleic acids research · 2017Review
- Silencing of HIV-1 by AgoshRNA molecules.Gene therapy · 2017Article
- Article
- Mutation of nucleotides around the +1 position of type 3 polymerase III promoters: The effect on transcriptional activity and start site usage.Transcription · 2017Article
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Recent evidence indicates that shRNAs with a relatively short basepaired stem do not require Dicer processing, but instead are processed by the Argonaute 2 protein (Ago2). We named these molecules AgoshRNAs as both their processing and silencing function are mediated by Ago2. This alternative processing yields only a single RNA guide strand, which can avoid off-target effects induced by the passenger strand of regular shRNAs. It is important to understand this alternative processing route in mechanistic detail such that one can design improved RNA reagents. We verified that AgoshRNAs trigger site-specific cleavage of a complementary mRNA. Second, we document the importance of the identity of the 5΄-terminal nucleotide and its basepairing status for AgoshRNA activity. AgoshRNA activity is significantly reduced or even abrogated with C or U at the 5΄-terminal and is enhanced by introduction of a bottom mismatch and 5΄-terminal nucleotide A or G. The 5΄-terminal RNA nucleotide also represents the +1 position of the transcriptional promoter in the DNA, thus further complicating the analysis. Indeed, we report that +1 modification affects the transcriptional efficiency and accuracy of start site selection, with A or G as optimal nucleotide. These combined results allow us to propose general rules for the design and expression of potent AgoshRNA molecules.
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