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

6.6K
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...
6.6K
Spontaneous and Induced Mutations01:30

Spontaneous and Induced Mutations

2.0K
Spontaneous mutations arise infrequently during DNA replication due to errors in the process. A key factor behind these errors is tautomeric shifts in nitrogenous bases, where bases transition from keto to enol forms or amino to imino forms. This shift can alter base-pairing rules, leading to mutations. Additionally, reactive oxygen species (ROS) arising from aerobic metabolism can damage DNA, resulting in depurination (loss of a purine base) or depyrimidination (loss of a pyrimidine base).
2.0K
Mismatch Repair01:20

Mismatch Repair

6.3K
Organisms are capable of detecting and fixing nucleotide mismatches that occur during DNA replication. This sophisticated process requires identifying the new strand and replacing the erroneous bases with correct nucleotides. Mismatch repair is coordinated by many proteins in both prokaryotes and eukaryotes.
The Mutator Protein Family Plays a Key Role in DNA Mismatch Repair
The human genome has more than 3 billion base pairs of DNA per cell. Prior to cell division, that vast amount of genetic...
6.3K
In-vitro Mutagenesis01:16

In-vitro Mutagenesis

16.0K
To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
16.0K
Targeted Cancer Therapies02:57

Targeted Cancer Therapies

8.6K
The targeted cancer therapies, also known as “molecular targeted therapies,” take advantage of the molecular and genetic differences between the cancer cells and the normal cells. It needs a thorough understanding of the cancer cells to develop drugs that can target specific molecular aspects that drive the growth, progression, and spread of cancer cells without affecting the growth and survival of other normal cells in the body.
There are several types of targeted therapies against...
8.6K

You might also read

Related Articles

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

Sort by
Same author

Discovery of a Two-Step Enzyme Cascade Converting Aspartate to Aminomalonate in Peptide Natural Product Biosynthesis.

Journal of the American Chemical Society·2025
Same author

Evolutionary Spread of Distinct O-methyltransferases Guides the Discovery of Unique Isoaspartate-Containing Peptides, Pamtides.

Advanced science (Weinheim, Baden-Wurttemberg, Germany)·2023
Same author

Exploring the Diverse Landscape of Biaryl-Containing Peptides Generated by Cytochrome P450 Macrocyclases.

Journal of the American Chemical Society·2023
Same author

Discovery and Biosynthesis of Cihunamides, Macrocyclic Antibacterial RiPPs with a Unique C-N Linkage Formed by CYP450 Catalysis.

Angewandte Chemie (International ed. in English)·2023
Same author

A dual gene-specific mutator system installs all transition mutations at similar frequencies in vivo.

Nucleic acids research·2023
Same author

Bioinformatic Expansion of Borosins Uncovers Trans-Acting Peptide Backbone <i>N</i>-Methyltransferases in Bacteria.

Biochemistry·2022

Related Experiment Video

Updated: Jan 10, 2026

Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast
08:48

Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast

Published on: January 26, 2017

16.3K

Continuous targeted hypermutation with a tunable mutation window.

Chanwoo Lee1, Seokhee Kim2,3

  • 1Department of Chemistry, Seoul National University, Seoul, Republic of Korea.

Nature Communications
|November 25, 2025
PubMed
Summary

Researchers developed RESPECTevo, a system for targeted DNA hypermutation in living cells. This tool enables rapid sequence diversification for continuous protein evolution in biotechnology.

More Related Videos

Following the Dynamics of Structural Variants in Experimentally Evolved Populations
04:52

Following the Dynamics of Structural Variants in Experimentally Evolved Populations

Published on: February 3, 2023

1.3K
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

7.7K

Related Experiment Videos

Last Updated: Jan 10, 2026

Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast
08:48

Targeted in Situ Mutagenesis of Histone Genes in Budding Yeast

Published on: January 26, 2017

16.3K
Following the Dynamics of Structural Variants in Experimentally Evolved Populations
04:52

Following the Dynamics of Structural Variants in Experimentally Evolved Populations

Published on: February 3, 2023

1.3K
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

7.7K

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Synthetic Biology

Background:

  • Targeted in vivo hypermutation allows for rapid DNA sequence diversification and biomolecule evolution.
  • Somatic hypermutation in antibody affinity maturation exemplifies this strategy, focusing on specific intragenic regions.
  • Current molecular tools for targeted hypermutation with adjustable range are limited.

Purpose of the Study:

  • To present RESPECTevo, a novel system for focused hypermutation of user-defined DNA regions.
  • To enable targeted hypermutation of endogenous and heterologous DNA sequences up to ~200 bp with precise boundaries.
  • To facilitate continuous protein evolution for broader biotechnological applications.

Main Methods:

  • Utilizing the Escherichia coli Cascade complex for its specific targeting, spacer length flexibility, and mismatch tolerance.
  • Implementing a system for focused hypermutation within predefined DNA boundaries.
  • Enabling modulation of the target range and continuous hypermutation across dual regions.

Main Results:

  • Demonstrated highly specific targeted hypermutation in user-defined regions.
  • Showcased the ability to modulate the target range of hypermutation.
  • Successfully achieved continuous hypermutation across dual DNA regions.
  • Established RESPECTevo as an efficient platform for hypermutating specific DNA targets in living cells.

Conclusions:

  • RESPECTevo provides an efficient platform for targeted DNA hypermutation in living cells.
  • The system facilitates continuous protein evolution for biotechnological advancements.
  • Enables precise control over hypermutation targets and range, expanding potential applications.