Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

Pre-mRNA Processing: Modification of pre-mRNA Ends01:35

Pre-mRNA Processing: Modification of pre-mRNA Ends

9.7K
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a cap to the 5' end of the growing transcript. In this process, a 5' phosphate is replaced by modified guanosine that has a methyl group attached (7-methyl guanosine). This 5' cap helps...
9.7K
pre-mRNA Processing02:01

pre-mRNA Processing

53.2K
In eukaryotic cells, transcripts made by RNA polymerase are modified and processed before exiting the nucleus. Unprocessed RNA is called precursor mRNA or pre-mRNA to distinguish it from mature mRNA.
Once about 20-40 ribonucleotides have been joined together by RNA polymerase, a group of enzymes adds a “cap” to the 5’ end of the growing transcript. In this process, a 5’ phosphate is replaced by modified guanosine that has a methyl group attached to it (7-Methyl...
53.2K
RNA Editing02:23

RNA Editing

9.1K
RNA editing is a post-transcriptional modification where a precursor mRNA (pre-mRNA) nucleotide sequence is changed by base insertion, deletion, or modification. The extent of RNA editing varies from a few hundred bases, in mitochondrial DNA of trypanosomes, to a just single base, in nuclear genes of mammals. Even a single base change in the pre-mRNA can convert a codon for one amino acid into the codon for another amino acid or a stop codon. This type of re-coding can significantly affect the...
9.1K
RNA Splicing01:32

RNA Splicing

56.7K
Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...
56.7K
Regulation of Expression at Multiple Steps01:23

Regulation of Expression at Multiple Steps

987
The gene expression in cells is regulated at different stages: (i) transcription, (ii) RNA processing, (iii) RNA localization, and (iv) translation. Transcriptional regulation is mediated by regulatory proteins such as transcription factors, activators, or repressors—these control gene expression by initiating or inhibiting the transcription of genes. Once a precursor or pre-mRNA is produced, it undergoes post-transcriptional modification, including 5' capping, splicing, and the...
987
Ribosomal RNA Synthesis02:53

Ribosomal RNA Synthesis

13.4K
Ribosome synthesis is a highly complex and coordinated process involving more than 200 assembly factors. The synthesis and processing of ribosomal components occurs not only in the nucleolus but also in the nucleoplasm and the cytoplasm of eukaryotic cells.
Ribosome biogenesis begins with the synthesis of 5S and 45S pre-rRNAs by distinct RNA polymerases. The primary transcripts are extensively processed and modified before they are bound and folded by ribosomal proteins and assembly factors,...
13.4K

You might also read

Related Articles

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

Sort by
Same author

Human RNA ligase 1 as a novel regulator of ribosome function and translation under oxidative stress.

Nucleic acids research·2026
Same author

ModiCal: A Targeted Calibration Workflow for Site-Specific m<sup>5</sup>C Validation by Nanopore Direct RNA Sequencing.

ACS chemical biology·2026
Same author

Real-time transcriptomic profiling in distinct experimental conditions.

eLife·2026
Same author

tRNA modification landscapes in streptococci: shared losses and clade-specific adaptations.

Open biology·2026
Same author

2'-O-Methylation maintains ribosome structural and translation integrity.

Molecular cell·2026
Same author

Mapping human pre-rRNA processing and modification at single nucleotide resolution using long read nanopore sequencing.

Nature communications·2026

Related Experiment Video

Updated: Aug 29, 2025

In Vitro Synthesis of Modified mRNA for Induction of Protein Expression in Human Cells
10:07

In Vitro Synthesis of Modified mRNA for Induction of Protein Expression in Human Cells

Published on: November 13, 2014

25.1K

Synthesis of point-modified mRNA.

Jasmin Hertler1, Kaouthar Slama1, Benedikt Schober1

  • 1Institute of Pharmaceutical and Biomedical Sciences, Johannes Gutenberg-Universität, Staudinger Weg 5, D-55128 Mainz, Germany.

Nucleic Acids Research
|September 5, 2022
PubMed
Summary

Researchers developed novel methods for creating site-specific modified messenger RNAs (mRNA) for therapeutic applications. These techniques enable precise internal mRNA modifications, improving translation and reducing immune responses for advanced vaccine and drug development.

More Related Videos

Using Lipid Nanoparticles for the Delivery of Chemically Modified mRNA into Mammalian Cells
10:02

Using Lipid Nanoparticles for the Delivery of Chemically Modified mRNA into Mammalian Cells

Published on: June 10, 2022

2.3K
Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism
11:37

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Published on: July 28, 2017

19.1K

Related Experiment Videos

Last Updated: Aug 29, 2025

In Vitro Synthesis of Modified mRNA for Induction of Protein Expression in Human Cells
10:07

In Vitro Synthesis of Modified mRNA for Induction of Protein Expression in Human Cells

Published on: November 13, 2014

25.1K
Using Lipid Nanoparticles for the Delivery of Chemically Modified mRNA into Mammalian Cells
10:02

Using Lipid Nanoparticles for the Delivery of Chemically Modified mRNA into Mammalian Cells

Published on: June 10, 2022

2.3K
Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism
11:37

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Published on: July 28, 2017

19.1K

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Biotechnology

Background:

  • Synthetic messenger RNA (mRNA) is crucial for therapeutics and vaccines.
  • Current mRNA modification methods (using modified triphosphate nucleotides) offer random or complete incorporation, limiting precise control.
  • Site-specific mRNA modification presents significant technical challenges.

Purpose of the Study:

  • To develop fundamental techniques for isolating long, translatable, and internally point-modified mRNAs.
  • To enable functional analysis of site-specific mRNA modifications, such as ribose methylations.

Main Methods:

  • Utilized three-way-one-pot splint ligations for mRNA construction.
  • Developed mRNA isolation via real-time elution from agarose gels using blue light visualization.
  • Quantified co-eluting agarose particles and assessed their impact on mRNA translation.
  • Measured EGFP protein translation using 35S-labelled methionine and in-gel fluorescence.

Main Results:

  • Successfully isolated long, translatable mRNAs with internal point modifications.
  • Blue light visualization enabled damage-free mRNA recovery from gels.
  • Isolated mRNA demonstrated efficient translation into functional EGFP protein in vitro.
  • Confirmed that co-eluting agarose particles did not impede mRNA translation.

Conclusions:

  • Established foundational techniques for producing site-specifically modified mRNAs.
  • The developed methods allow for precise control over mRNA modifications, enhancing therapeutic potential.
  • This work facilitates the functional analysis of specific mRNA modifications for improved drug and vaccine design.