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

Proofreading01:31

Proofreading

6.3K
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...
6.3K
Translesion DNA Polymerases02:10

Translesion DNA Polymerases

10.0K
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...
10.0K
PCR01:32

PCR

207.3K
Overview
207.3K
Bacterial RNA Polymerase00:43

Bacterial RNA Polymerase

29.6K
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...
29.6K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.0K
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...
6.0K
The Replisome03:01

The Replisome

33.6K
DNA replication is carried out by a large complex of proteins that act in a coordinated matter to achieve high-fidelity DNA replication. Together this complex is known as the DNA replication machinery or the replisome.
The synthesis of the leading and lagging strands is a highly coordinated process. To explain this, the “Trombone model” was proposed by Bruce Alberts in 1980. The DNA loop formation starts when a primer is synthesized on the parent lagging strand. The loop grows with...
33.6K

You might also read

Related Articles

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

Sort by
Same author

DNA synthesis: When amino acids act as a template.

Current biology : CB·2026
Same author

How custom polymerases are driving innovation in synthetic biology.

Current opinion in chemical biology·2026
Same author

Xeno-nucleic acids support formation of Ag(I)-mediated duplexes and silver nanoclusters.

Nucleic acids research·2026
Same author

Allele-specific knockdown by an engineered DNAzyme capable of RNase H1 evasion.

Nucleic acids research·2026
Same author

Rapid evolution of a highly efficient RNA polymerase by homologous recombination.

Nature chemical biology·2026
Same author

Directed evolution of a TNA polymerase identifies independent paths to fidelity and catalysis.

Nature communications·2025

Related Experiment Video

Updated: Jul 12, 2025

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

14.2K

Engineering TNA polymerases through iterative cycles of directed evolution.

Eric J Yik1, Victoria A Maola1, John C Chaput2

  • 1Department of Pharmaceutical Sciences, University of California, Irvine, CA, United States.

Methods in Enzymology
|November 1, 2023
PubMed
Summary

Researchers engineered DNA polymerases to synthesize artificial genetic polymers called threose nucleic acids (TNA). This directed evolution strategy enables copying genetic information between DNA and XNA, advancing biotechnology.

Keywords:
EvolutionPolymerase engineeringThreose nucleic acid (TNA)XNAXeno-nucleic acid

More Related Videos

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

7.3K
A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes
13:30

A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes

Published on: November 7, 2012

18.1K

Related Experiment Videos

Last Updated: Jul 12, 2025

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

14.2K
In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity
09:16

In Vitro Directed Evolution of a Restriction Endonuclease with More Stringent Specificity

Published on: March 25, 2020

7.3K
A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes
13:30

A New Screening Method for the Directed Evolution of Thermostable Bacteriolytic Enzymes

Published on: November 7, 2012

18.1K

Area of Science:

  • Biotechnology
  • Synthetic Biology
  • Chemical Biology
  • Molecular Biology

Background:

  • DNA polymerases are essential for genetic information manipulation.
  • Natural polymerases cannot process artificial genetic polymers (xeno-nucleic acids or XNAs) due to unique backbone structures.
  • Directed evolution can create engineered polymerases capable of interacting with XNAs.

Purpose of the Study:

  • To develop a directed evolution strategy for discovering polymerases that synthesize threose nucleic acid (TNA) using DNA templates.
  • To demonstrate a generalizable method for engineering DNA-modifying enzymes.

Main Methods:

  • Employed a directed evolution workflow.
  • Utilized library generation and expression in E. coli.
  • Implemented high-throughput microfluidics-based screening in water-in-oil droplets.
  • Incorporated plasmid recovery, secondary screening, and library regeneration.

Main Results:

  • Successfully discovered polymerases capable of synthesizing TNA on DNA templates.
  • Established a robust workflow for directed evolution of DNA polymerases.
  • Demonstrated the potential for engineering polymerases to recognize and process XNAs.

Conclusions:

  • The developed directed evolution strategy is effective for discovering polymerases that synthesize TNA.
  • This approach is broadly applicable to engineering DNA-modifying enzymes for various biotechnological applications.
  • Advances in XNA technology are facilitated by the ability to synthesize and process these artificial nucleic acids.