ArticleNature protocols2009
Investigation of mRNA quadruplex formation in Escherichia coli.
Article in Nature protocols, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 14 papers.
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Who cites it
14 citing papers in PubMed, 38 citations in OpenAlex.
- RNA G-quadruplexes (rG4s): genomics and biological functions.Nucleic acids research · 2021Review
- Mapping and characterization of G-quadruplexes in the genome of the social amoeba Dictyostelium discoideum.Nucleic acids research · 2019Article
- Surface Plasmon Resonance kinetic analysis of the interaction between G-quadruplex nucleic acids and an anti-G-quadruplex monoclonal antibody.Biochimica et biophysica acta. General subjects · 2018Article
- G-Quadruplex Identification in the Genome of Protozoan Parasites Points to Naphthalene Diimide Ligands as New Antiparasitic Agents.Journal of medicinal chemistry · 2018Article
- Article
- Mms1 binds to G-rich regions in Saccharomyces cerevisiae and influences replication and genome stability.Nucleic acids research · 2017Article
- The cellular protein nucleolin preferentially binds long-looped G-quadruplex nucleic acids.Biochimica et biophysica acta. General subjects · 2017Article
- The Herpes Simplex Virus-1 genome contains multiple clusters of repeated G-quadruplex: Implications for the antiviral activity of a G-quadruplex ligand.Antiviral research · 2015Article
- A variable homopolymeric G-repeat defines small RNA-mediated posttranscriptional regulation of a chemotaxis receptor in Helicobacter pylori.Proceedings of the National Academy of Sciences of the United States of America · 2014Article
- A dynamic G-quadruplex region regulates the HIV-1 long terminal repeat promoter.Journal of medicinal chemistry · 2013Article
- Thermozymes: Synthetic RNA thermometers based on ribozyme activity.RNA biology · 2013Article
- Formation of a unique cluster of G-quadruplex structures in the HIV-1 Nef coding region: implications for antiviral activity.PloS one · 2013Article
- Twins, quadruplexes, and more: functional aspects of native and engineered RNA self-assemblyFrontiers in life science · 2012Article
- Predictable suppression of gene expression by 5'-UTR-based RNA quadruplexes.Nucleic acids research · 2009Article
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The protocol presented here allows for the investigation of the formation of unusual nucleic acid structures in the 5'-untranslated region (UTR) of bacteria by correlating gene expression levels to the in vitro stability of the respective structure. In particular, we describe the introduction of G-quadruplex forming sequences close to the ribosome-binding site (RBS) on the mRNA of a reporter gene and the subsequent read-out of the expression levels. Insertion of a stable secondary structure results in the cloaking of RBS and eventually reduced gene expression levels. The structures and stability of the introduced sequences are further characterized by circular dichroism (CD) spectroscopy and thermal melting experiments. The extent of inhibition is then correlated to the stability of the respective quadruplex structure, allowing judgement of whether factors other than thermodynamic stability affect the formation of a given quadruplex sequence in vivo. Measuring gene expression levels takes 2 d including cloning; CD experiments take 5 hours per experiment.
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Registered trials
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.