Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

Riboswitches

Riboswitches

Riboswitches are non-coding mRNA domains that regulate the transcription and translation of downstream genes without the help of proteins. Riboswitches bind directly to a metabolite and can form unique stem-loop or hairpin structures in response to the amount of the metabolite present. They have two distinct regions – a metabolite-binding aptamer and an expression platform.
The aptamer has high specificity for a particular metabolite which allows riboswitches to specifically regulate...

Ribosome Profiling

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...

Types of RNA

Types of RNA

Overview
Three main types of RNA are involved in protein synthesis: messenger RNA (mRNA), transfer RNA (tRNA), and ribosomal RNA (rRNA). These RNAs perform diverse functions and can be broadly classified as protein-coding or non-coding RNA. Non-coding RNAs play important roles in the regulation of gene expression in response to developmental and environmental changes. Non-coding RNAs in prokaryotes can be manipulated to develop more effective antibacterial drugs for human or animal use.
RNA...

Transcriptional Regulation: Riboswitches

Transcriptional Regulation: Riboswitches

Riboswitches are RNA elements that regulate gene expression by altering their secondary structures in response to specific effector molecules. These elements, located in the leader regions of certain mRNAs, act as transcriptional regulators by toggling between alternative conformations to control downstream gene expression. Riboswitch-mediated regulation is a precise mechanism for modulating biosynthetic pathways, as exemplified by the riboflavin biosynthesis pathway in Bacillus...

Applications Of NMR In Biology

Applications Of NMR In Biology

Nuclear magnetic resonance (NMR) spectroscopy is a very valuable analytical technique for researchers. It has been used for more than 50 years as an analytical tool. F. Bloch and E. Purcell formulated NMR in 1946 and won the 1952 Nobel Prize in Physics for their work. Biological macromolecules such as proteins, nucleic acids, lipids, and organic molecules including pharmaceutical compounds, can be studied using this versatile tool that exploits the magnetic properties of certain nuclei.

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by

Same author

Correction: <i>In situ</i> formation of transcriptional modulators using non-canonical DNA i-motifs.

Chemical science·2026

Same author

Integrated NMR/MD investigation reveals differences after reweighting in conformational ensembles of GAAG and GCAA tetraloops.

RNA (New York, N.Y.)·2026

Same author

SLX4IP limits replication stress globally and at ALT telomeres.

The EMBO journal·2026

Same author

RNA•DNA:DNA Triplex Formation Modulates Individual Base Pair Stabilities in the DNA Target Duplex.

RNA (New York, N.Y.)·2026

Same author

DARPins as pan-reactivators of temperature-sensitive p53 cancer mutants.

Proceedings of the National Academy of Sciences of the United States of America·2026

Same author

Structure and sequence characteristics of 5'-stem-loop 1 modulate the escape from nsp1-mediated repression in SARS-CoV-2 variants.

Nucleic acids research·2026

Search research articles

Related Experiment Video

Updated: Dec 7, 2025

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases

Published on: June 30, 2019

In-Cell NMR Spectroscopy of Functional Riboswitch Aptamers in Eukaryotic Cells.

P Broft¹, S Dzatko^2,3, M Krafcikova^2,4

¹Center for Biomolecular Magnetic Resonance (BMRZ), Institute for Organic Chemistry and Chemical Biology, Goethe University, Max-von-Laue-Str. 7, 60438, Frankfurt/M., Germany.

Angewandte Chemie (International Ed. in English)

|September 25, 2020

Summary

This summary is machine-generated.

This study demonstrates in-cell NMR spectroscopy observing 2'-deoxyguanosine binding to a riboswitch in Xenopus oocytes and HeLa cells. The findings confirm conserved binding modes and enable studying larger, non-modified RNA within cells.

Keywords:

2′-deoxyguanosine riboswitch HeLa cells RNA structures aptamers structural biology

More Related Videos

An Assay for Quantifying Protein-RNA Binding in Bacteria

An Assay for Quantifying Protein-RNA Binding in Bacteria

Published on: June 12, 2019

Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor

Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor

Published on: March 9, 2021

Related Experiment Videos

Last Updated: Dec 7, 2025

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases

NMR-Based Activity Assays for Determining Compound Inhibition, IC50 Values, Artifactual Activity, and Whole-Cell Activity of Nucleoside Ribohydrolases

Published on: June 30, 2019

An Assay for Quantifying Protein-RNA Binding in Bacteria

An Assay for Quantifying Protein-RNA Binding in Bacteria

Published on: June 12, 2019

Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor

Monitoring Protein-Ligand Interactions in Human Cells by Real-Time Quantitative In-Cell NMR using a High Cell Density Bioreactor

Published on: March 9, 2021

Area of Science:

Biochemistry
Molecular Biology
Cell Biology

Background:

Riboswitches are RNA molecules that regulate gene expression in response to small molecule binding.
Bacterial riboswitches have been extensively studied in vitro, but their behavior in eukaryotic cellular environments remains less understood.
In-cell NMR spectroscopy offers a powerful tool to study biomolecular interactions within living cells.

Purpose of the Study:

To observe the binding of 2 -deoxyguanosine to its cognate riboswitch aptamer domain within eukaryotic cells.
To determine if the binding mode of a prokaryotic riboswitch is conserved in a eukaryotic cellular environment.
To establish and validate a methodological setup for in-cell NMR studies of larger, non-modified RNA molecules in mammalian cells.

Main Methods:

In-cell NMR spectroscopy was performed on Xenopus laevis oocytes and human HeLa cells.
The aptamer domain of the 2 -deoxyguanosine-sensing riboswitch was introduced into these eukaryotic cells.
The binding of 2 -deoxyguanosine to the riboswitch was monitored using NMR signals.

Main Results:

The binding of 2 -deoxyguanosine to the riboswitch was successfully observed in both Xenopus oocytes and HeLa cells.
The binding mode of the riboswitch in the cellular environment was consistent with previously established in vitro findings.
The study demonstrated the feasibility of using in-cell NMR for characterizing larger (≈70 nt), chemically non-modified RNA molecules in mammalian cells, overcoming previous limitations with shorter, modified fragments.

Conclusions:

The aptamer domain of the 2 -deoxyguanosine riboswitch maintains its ligand-binding properties and conserved binding mode within eukaryotic cells.
In-cell NMR spectroscopy is a viable technique for studying the structure and function of larger, functional RNA molecules in their native cellular context.
This work expands the scope of in-cell NMR applications for RNA research in mammalian systems.