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

Homologous Recombination02:31

Homologous Recombination

50.8K
The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
50.8K
Gene Conversion02:08

Gene Conversion

9.9K
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...
9.9K
Conservative Site-specific Recombination and Phase Variation02:53

Conservative Site-specific Recombination and Phase Variation

6.1K
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...
6.1K
Base Excision Repair01:54

Base Excision Repair

22.8K
One of the common DNA damages is the chemical alteration of single bases by alkylation, oxidation, or deamination. The altered bases cause mispairing and strand breakage during replication. This type of damage causes minimal change to the DNA double helix structure and can be repaired by the base excision repair (BER) pathways. BER corrects damaged DNA sequences by removing the damaged base and restoring the original base sequence using the complementary strand as a template.
The first step of...
22.8K
Proofreading01:31

Proofreading

6.5K
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...
6.5K

You might also read

Related Articles

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

Sort by
Same author

Yeast Molecular Display for the Screening and Production of Immunological Proteins Using Nanobody as an Example.

Methods in molecular biology (Clifton, N.J.)·2026
Same author

Autonomous ribosome biogenesis in vitro.

Nature communications·2025
Same author

Bacterial removal using liposomes and an anionic adsorber.

Journal of bioscience and bioengineering·2024
Same author

In vitro selection of dye-fluorescence-enhancing peptide aptamer by cDNA display.

Analytical biochemistry·2024
Same author

Secretion of mitochondrial DNA via exosomes promotes inflammation in Behçet's syndrome.

The EMBO journal·2023
Same author

Application of peptide barcoding to obtain high-affinity anti-PD-1 nanobodies.

Journal of bioscience and bioengineering·2023

Related Experiment Video

Updated: Aug 22, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
09:51

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

34.2K

Improving Precise Genome Editing Using Donor DNA/gRNA Hybrid Duplex Generated by Complementary Bases.

Wataru Aiba1, Takamitsu Amai1, Mitsuyoshi Ueda1

  • 1Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan.

Biomolecules
|November 11, 2022
PubMed
Summary
This summary is machine-generated.

Researchers improved CRISPR genome editing efficiency by combining guide RNA and donor DNA into a single hybrid molecule. This DGybrid approach enhances homology-directed repair (HDR) for more precise genetic modifications.

Keywords:
CRISPR/Cas9DNA/RNA hybridSaccharomyces cerevisiaegenome editingguide RNAsingle-stranded oligodeoxynucleotide

More Related Videos

Genome Editing in Mammalian Cell Lines using CRISPR-Cas
07:56

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

22.0K
CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
08:22

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

Published on: March 12, 2018

15.0K

Related Experiment Videos

Last Updated: Aug 22, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
09:51

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

34.2K
Genome Editing in Mammalian Cell Lines using CRISPR-Cas
07:56

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

22.0K
CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy
08:22

CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy

Published on: March 12, 2018

15.0K

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR/Cas9 enables precise genome editing via DNA double-strand breaks (DSBs).
  • Homology-directed repair (HDR) uses donor DNA for precise editing but is often inefficient.
  • Low HDR efficiency limits the widespread application of CRISPR-based genome editing.

Purpose of the Study:

  • To develop a novel donor DNA/guide RNA (gRNA) hybrid duplex (DGybrid) to enhance HDR efficiency.
  • To improve the concentration of donor DNA at the DNA cleavage site.
  • To create a universal and concise tool for efficient genome editing.

Main Methods:

  • Constructed a DGybrid by combining a sequence-extended gRNA with a single-stranded oligodeoxynucleotide (ssODN).
  • Evaluated DGybrid efficiency in *Saccharomyces cerevisiae* for HDR-mediated genome editing.
  • Assessed nucleic acid introduction efficiency using flow cytometry.

Main Results:

  • Achieved a 1.8-fold improvement in HDR-mediated editing efficiency (from 35% to 62%) compared to separate introduction of gRNA and donor DNA.
  • Demonstrated efficient co-incorporation of RNA and ssODNs into cells via the DNA/RNA hybrid.
  • Confirmed the effectiveness of the DGybrid in enhancing genome editing outcomes.

Conclusions:

  • The DGybrid technique significantly boosts HDR efficiency in CRISPR/Cas9 genome editing.
  • This method facilitates simultaneous delivery and cellular uptake of essential editing components.
  • DGybrid presents a promising, universally applicable tool for advancing precise genome engineering.