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

Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

5.9K
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
5.9K
Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

28.5K
Unlike eukaryotes, bacteria use a single RNA Polymerase (RNAP) to transcribe all genes. The different subunits of bacterial RNAPhave distinct functions. The multisubunit structure of the bacterial RNAP helps the enzyme to maintain catalytic function, facilitate assembly, interact with DNA and RNA, and self-regulate its activity.
In most genes, the transcription site is a single base present upstream of the coding sequence. Though RNAP is a catalytically efficient enzyme, it does not recognize...
28.5K
Proofreading01:31

Proofreading

6.2K
Synthesis of new DNA molecules is carried out by the enzyme DNA polymerase, which adds nucleotides on the daughter strand complementary to the template DNA strand. DNA polymerase has a higher affinity to add the correct base and ensures fidelity during DNA replication. Furthermore,  it exhibits proofreading activity during replication, using an exonuclease domain that cuts off incorrect nucleotides from the nascent DNA strand.
Errors During Replication are Corrected by the DNA Polymerase...
6.2K
RNA Editing02:23

RNA Editing

8.9K
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...
8.9K
Translesion DNA Polymerases02:10

Translesion DNA Polymerases

9.8K
Translesion (TLS) polymerases rescue stalled DNA polymerases at sites of damaged bases by replacing the replicative polymerase and installing a nucleotide across the damaged site. Doing so, TLS allows additional time for the cell to repair the damage before resuming regular DNA replication.
TLS polymerases are found in all three domains of life - archaea, bacteria, and eukaryotes. Of the different classes of TLS polymerases, members of the Y family are fitted with specialized structures that...
9.8K
Eukaryotic RNA Polymerases00:58

Eukaryotic RNA Polymerases

23.3K
RNA Polymerase (RNAP) is conserved in all animals, with bacterial, archaeal, and eukaryotic RNAPs sharing significant sequence, structural, and functional similarities. Among the three eukaryotic RNAPs, RNA Polymerase II is most similar to bacterial RNAP in terms of both structural organization and folding topologies of the enzyme subunits. However, these similarities are not reflected in their mechanism of action.
All three eukaryotic RNAPs require specific transcription factors, of which the...
23.3K

You might also read

Related Articles

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

Sort by
Same author

Quantitative CRACI reveals transcriptome-wide distribution of RNA dihydrouridine at base resolution.

Nature communications·2025
Same author

Structural dynamics-guided engineering of a riboswitch RNA for evolving c-di-AMP synthases.

Science advances·2025
Same author

Author Correction: Engineering a DNA polymerase for modifying large RNA at specific positions.

Nature chemistry·2025
Same author

Ligand response of guanidine-IV riboswitch at single-molecule level.

eLife·2024
Same author

Divergent molecular assembly and catalytic mechanisms between bacterial and archaeal RNase P in pre-tRNA cleavage.

Proceedings of the National Academy of Sciences of the United States of America·2024
Same author

Occurrence, migration and health risks of fluorescent whitening agents and phthalates in bottled water.

Journal of hazardous materials·2024

Related Experiment Video

Updated: Jun 2, 2025

DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation
09:26

DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation

Published on: December 29, 2021

4.1K

Engineering a DNA polymerase for modifying large RNA at specific positions.

Dian Chen1, Zhanghui Han1, Xiaoge Liang1

  • 1State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.

Nature Chemistry
|January 13, 2025
PubMed
Summary

Scientists engineered a DNA polymerase variant for precise RNA modification. This method efficiently introduces diverse chemical modifications into RNA, enhancing its stability and protein production for research and therapeutics.

More Related Videos

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
07:38

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis

Published on: October 6, 2017

14.1K
DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
09:12

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation

Published on: September 16, 2019

8.2K

Related Experiment Videos

Last Updated: Jun 2, 2025

DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation
09:26

DNA-Tethered RNA Polymerase for Programmable In vitro Transcription and Molecular Computation

Published on: December 29, 2021

4.1K
DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis
07:38

DNA Polymerase Activity Assay Using Near-infrared Fluorescent Labeled DNA Visualized by Acrylamide Gel Electrophoresis

Published on: October 6, 2017

14.1K
DNAzyme-dependent Analysis of rRNA 2’-O-Methylation
09:12

DNAzyme-dependent Analysis of rRNA 2’-O-Methylation

Published on: September 16, 2019

8.2K

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Synthetic Biology

Background:

  • Precise synthesis of modified large RNA is crucial for research and therapeutics but faces limitations in current methods.
  • Engineered DNA polymerases offer advantages over traditional RNA polymerases for RNA labeling and modification.

Purpose of the Study:

  • To engineer a DNA polymerase variant for precise incorporation of diverse modifications into RNA.
  • To demonstrate the efficiency and versatility of this engineered polymerase for RNA functionalization.

Main Methods:

  • Semi-rational design of DNA polymerase variants.
  • Site-specific incorporation of base, 2'-ribose, and backbone modifications into RNA.
  • Assessment of modification efficiency and impact on messenger RNA (mRNA) stability and protein production.

Main Results:

  • Engineered DNA polymerase achieved >85% efficiency for most modifications.
  • Successfully introduced 2'-O-methyl, phosphorothioate, N4-acetylcytidine, and fluorophore modifications at specific sites.
  • Modified mRNA demonstrated enhanced stability and altered protein production.

Conclusions:

  • The engineered DNA polymerase provides a versatile and efficient tool for comprehensive RNA functionalization.
  • This method enables precise introduction of diverse modifications, irrespective of RNA length and sequence.
  • The developed technique holds promise for advancing RNA-based research and therapeutic applications.