Search research articles

ABOUT JoVE

Overview Leadership Blog JoVE Help Center

AUTHORS

Publishing Process Editorial Board Scope & Policies Peer Review FAQ Submit

LIBRARIANS

Testimonials Subscriptions Access Resources Library Advisory Board FAQ

RESEARCH

JoVE Journal Methods Collections JoVE Encyclopedia of Experiments Archive

EDUCATION

JoVE Core JoVE Business JoVE Science Education JoVE Lab Manual Faculty Resource Center Faculty Site

Terms & Conditions of Use

Related Concept Videos

CRISPR/Cas9 Genome Editing

CRISPR/Cas9 Genome Editing

The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...

In-vitro Mutagenesis

In-vitro Mutagenesis

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.

You might also read

Related Articles

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

Sort by

Same author

Portal Vein Thrombosis in Pediatric Liver Transplantation With Technical Variant Grafts: An International Multicenter Analysis of Risk and Prognostic Factors.

Transplantation·2026

Same author

Can AI simplify the alphabet of life?

Science (New York, N.Y.)·2026

Same author

DNA assembly with Sidewinder via 3-way junctions.

Nature reviews. Genetics·2026

Same author

Temperature and refractive index sensor based on a spherical-structure Mach-Zehnder interferometer combined with a long-period fiber grating.

Applied optics·2026

Same author

Construction of complex and diverse DNA sequences using DNA three-way junctions.

Nature·2026

Same author

Recruiting ESCRT to single-chain heterotrimer peptide MHCI releases antigen-presenting vesicles that stimulate T cells selectively.

Proceedings of the National Academy of Sciences of the United States of America·2025

Same journal

Cap 2'-O-methyltransferase CMTR2 regulates male meiosis independent of its methyltransferase activity.

Nucleic acids research·2026

Same journal

APE1 binds and processes abasic sites present in i-motif DNA and cooperates with PCBP1 in maintenance of telomeric stability.

Nucleic acids research·2026

Same journal

Acquisition of a novel restriction modification system regulates genetic flux and gene expression in the hypervirulent and globally disseminated CC17 lineage of group B Streptococcus.

Nucleic acids research·2026

Same journal

Trans-species microRNAs from the parasitic plant Cuscuta campestris specifically avoid loading onto self Argonautes.

Nucleic acids research·2026

Same journal

Neurochondrin promotes U5 snRNP maturation by regulating AAR2 release from PRPF8.

Nucleic acids research·2026

Same journal

Elongationless start-stop elements are stress-resilient translation gates that are more repressive than uTranslons.

Nucleic acids research·2026

See all related articles

Search research articles

Related Experiment Video

Updated: Aug 6, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

Genome manipulation by guide-directed Argonaute cleavage.

Shan Huang¹, Kaihang Wang², Stephen L Mayo^1,2

¹Division of Chemistry and Chemical Engineering, California Institute of Technology, MC 114-96, 1200 East California Boulevard, Pasadena, CA 91125, USA.

Nucleic Acids Research

|March 17, 2023

Summary

This summary is machine-generated.

Prokaryotic Argonautes (pAgos) can be mutagenic, inducing DNA cleavage and recombination in bacterial genomes. This guide-directed DNA interference has potential applications in genetic engineering and pAgo characterization.

More Related Videos

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

Published on: April 30, 2018

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

Related Experiment Videos

Last Updated: Aug 6, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

A Rapid and Facile Pipeline for Generating Genomic Point Mutants in C. elegans Using CRISPR/Cas9 Ribonucleoproteins

Published on: April 30, 2018

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

Area of Science:

Molecular Biology
Genetics
Microbiology

Background:

Prokaryotic Argonautes (pAgos) are known to mediate DNA interference using guide DNA to cleave target DNA.
Clostridium butyricum Ago (CbAgo) induces DNA interference and double-stranded breaks (DSBs) in target DNA, aiding host defense against foreign DNA.
The mutagenic potential of CbAgo-mediated DNA cleavage remained uninvestigated.

Purpose of the Study:

To investigate whether CbAgo-mediated DNA cleavage is mutagenic.
To explore the mechanisms underlying CbAgo-induced DNA cleavage and recombination in Escherichia coli.
To assess the utility of CbAgo's mutagenic activity in genetic manipulation.

Main Methods:

Utilized plasmid-encoded guide sequences to direct CbAgo cleavage of target sites in the E. coli genome.
Analyzed chromosome recombination between homologous sequences induced by CbAgo cleavage.
Studied the role of DSBs and RecBCD processing in the recombination mechanism.
Investigated the impact of CbAgo cleavage on cell growth in RecA-deficient E. coli strains.

Main Results:

CbAgo-directed cleavage of genomic target sites induced chromosome recombination between homologous sequences in E. coli.
Recombination rates correlated with CbAgo DNA cleavage activity.
Mechanistic studies indicated that DSBs and RecBCD processing are involved in the recombination process.
Guide-directed CbAgo cleavage on chromosomes severely impaired cell growth in RecA-deficient E. coli, enabling counter-selection for Lambda-Red recombineering.

Conclusions:

Guide-directed CbAgo cleavage of the host genome is mutagenic, leading to outcomes dependent on host DNA repair pathways.
This DNA-guided interference mechanism offers potential for broader genetic manipulation applications.
The study provides an in vivo assay for characterizing and engineering pAgo DNA cleavage activity.