ArticlePloS one2014
Reporters transiently transfected into mammalian cells are highly sensitive to translational repression induced by dsRNA expression.
Article in PloS one, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 33 citations in OpenAlex.
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- Functional canonical RNAi in mice expressing a truncated Dicer isoform and long dsRNA.EMBO reports · 2024Article
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- Maladaptation after a virus host switch leads to increased activation of the pro-inflammatory NF-κB pathway.Proceedings of the National Academy of Sciences of the United States of America · 2022Article
- Inhibition of the integrated stress response by viral proteins that block p-eIF2-eIF2B association.Nature microbiology · 2020Article
- Potentials and pitfalls of transient in vitro reporter bioassays: interference by vector geometry and cytotoxicity in recombinant zebrafish cell lines.Archives of toxicology · 2020Article
- STARR-seq and UMI-STARR-seq: Assessing Enhancer Activities for Genome-Wide-, High-, and Low-Complexity Candidate Libraries.Current protocols in molecular biology · 2019Article
- Main constraints for RNAi induced by expressed long dsRNA in mouse cells.Life science alliance · 2019Article
- Resolving systematic errors in widely used enhancer activity assays in human cells.Nature methods · 2018Article
- Design, simplified cloning, andMolecular biology research communications · 2016Article
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In mammals, double-stranded RNA (dsRNA) can mediate sequence-specific RNA interference, activate sequence-independent interferon response, or undergo RNA editing by adenosine deaminases. We showed that long hairpin dsRNA expression had negligible effects on mammalian somatic cells--expressed dsRNA was slightly edited, poorly processed into siRNAs, and it did not activate the interferon response. At the same time, we noticed reduced reporter expression in transient co-transfections, which was presumably induced by expressed dsRNA. Since transient co-transfections are frequently used for studying gene function, we systematically explored the role of expressed dsRNA in this silencing phenomenon. We demonstrate that dsRNA expressed from transiently transfected plasmids strongly inhibits the expression of co-transfected reporter plasmids but not the expression of endogenous genes or reporters stably integrated in the genome. The inhibition is concentration-dependent, it is found in different cell types, and it is independent of transfection method and dsRNA sequence. The inhibition occurs at the level of translation and involves protein kinase R, which binds the expressed dsRNA. Thus, dsRNA expression represents a hidden danger in transient transfection experiments and must be taken into account during interpretation of experimental results.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.