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

In-vitro Mutagenesis01:16

In-vitro Mutagenesis

14.0K
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.
14.0K
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
Gene Conversion02:08

Gene Conversion

9.8K
Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
9.8K
Mismatch Repair01:20

Mismatch Repair

4.9K
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...
4.9K

You might also read

Related Articles

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

Sort by
Same author

Insertion orientation within the cassette affects gene-targeting success during ends-out recombination in the yeast Saccharomyces cerevisiae.

Current genetics·2022
See all related articles

Related Experiment Video

Updated: Jul 14, 2025

CIRCLE-Seq for Interrogation of Off-Target Gene Editing
08:23

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

675

Unilateral ends-out gene targeting increases mistargeting through supporting extensive single-strand assimilation.

Petar Tomev Mitrikeski1,2

  • 1Laboratory of Evolutionary Genetics, Division of Molecular Biology, Ruđer Bošković Institute, Zagreb, Croatia.

Yeast (Chichester, England)
|October 9, 2023
PubMed
Summary

This study reveals a unified mechanism for ends-out gene targeting in yeast, involving two pathways that initiate homologous invasion differently. Bilateral targeting appears to reduce errors by limiting strand assimilation.

Keywords:
bilateral homologous invasionends-out gene targetingrecombination mechanismsstrand assimilationunilateral homologous invasionyeast Saccharomyces cerevisiae

More Related Videos

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing
10:44

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

Published on: May 5, 2023

1.5K
In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
08:54

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression

Published on: March 29, 2019

7.1K

Related Experiment Videos

Last Updated: Jul 14, 2025

CIRCLE-Seq for Interrogation of Off-Target Gene Editing
08:23

CIRCLE-Seq for Interrogation of Off-Target Gene Editing

Published on: November 1, 2024

675
In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing
10:44

In Vitro Selection of Engineered Transcriptional Repressors for Targeted Epigenetic Silencing

Published on: May 5, 2023

1.5K
In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression
08:54

In vivo Application of the REMOTE-control System for the Manipulation of Endogenous Gene Expression

Published on: March 29, 2019

7.1K

Area of Science:

  • Molecular Biology
  • Genetics
  • Yeast as a model organism

Background:

  • Ends-out gene targeting, a method for allele replacement via homologous recombination, has significant fundamental and applied implications.
  • Understanding the precise mechanisms of homologous recombination in gene targeting is crucial for its effective application.

Purpose of the Study:

  • To investigate the mechanisms underlying ends-out gene targeting in Saccharomyces cerevisiae.
  • To differentiate between unilateral and bilateral targeting pathways and their respective frequencies.
  • To elucidate the role of strand assimilation in targeting accuracy and mistargeting.

Main Methods:

  • Development of an experimental system in yeast to quantify unilateral and bilateral targeting outcomes.
  • Quantitative analysis of genetic outcomes to infer targeting pathway probabilities.
  • Comparative analysis across different experimental setups.
  • Comprehensive qualitative analysis to reveal the underlying gene targeting mechanism.

Main Results:

  • Quantitative analysis initially indicated predominant bilateral targeting.
  • A subsequent analysis suggested a prevalence of unilateral targeting, raising questions about experimental artifacts.
  • A unified model of ends-out gene targeting involving two intertwined pathways was proposed.
  • Bilateral targeting was suggested to minimize mistargeting by limiting strand assimilation.

Conclusions:

  • Ends-out gene targeting operates via a single fundamental mechanism with two distinct pathways for homologous invasion initiation.
  • Bilateral targeting may enhance accuracy by restricting strand assimilation, contrasting with unilateral targeting's potential for extensive assimilation and mistargeting.
  • The findings provide a deeper understanding of homologous recombination and gene targeting mechanisms.