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

Restriction Enzymes

Restriction Enzymes

Restriction enzymes are bacterial enzymes used to cut DNA in a sequence-specific manner. To cleave DNA, they bind to specific palindromic sequences called restriction sites. Such palindromic DNA sequences or inverted repeats are commonly found in regions of functional significance, such as the origin of replication, gene operator sites, and regions containing transcription termination signals.
The host bacteria protect their own genomic DNA from these enzymes by methylating these sites. Some...

Maxam-Gilbert Sequencing

Maxam-Gilbert Sequencing

In the same year as the discovery of the Sanger sequencing method, another group of scientists, Allan Maxam and Walter Gilbert, demonstrated their chemical-cleavage method for DNA sequencing. The Maxam-Gilbert method relies on using different chemicals that can cleave the DNA sequence at specific sites, the separation of resulting DNA fragments of variable size using electrophoresis, and deciphering the DNA sequence from the resulting gel bands.
Challenges of the Maxam-Gilbert Method
The...

Mismatch Repair

Mismatch Repair

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...

Nucleotide Excision Repair

Nucleotide Excision Repair

Fixing Double-strand Breaks

Fixing Double-strand Breaks

The double-stranded structure of DNA has two major advantages. First, it serves as a safe repository of genetic information where one strand serves as the back-up in case the other strand is damaged. Second, the double-helical structure can be wrapped around proteins called histones to form nucleosomes, which can then be tightly wound to form chromosomes. This way, DNA chains up to 2 inches long can be contained within microscopic structures in a cell. A double-stranded break not only damages...

You might also read

Related Articles

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

Sort by

Same author

CASP11 refinement experiments with ROSETTA.

Proteins·2015

Same author

Structure of a designed tetrahedral protein assembly variant engineered to have improved soluble expression.

Protein science : a publication of the Protein Society·2015

Same author

Unique double-ring structure of the peroxisomal Pex1/Pex6 ATPase complex revealed by cryo-electron microscopy.

Proceedings of the National Academy of Sciences of the United States of America·2015

Same author

Mechanistic Analysis of an Engineered Enzyme that Catalyzes the Formose Reaction.

Chembiochem : a European journal of chemical biology·2015

Same author

Design of ordered two-dimensional arrays mediated by noncovalent protein-protein interfaces.

Science (New York, N.Y.)·2015

Same author

Designing Two-Dimensional Protein Arrays through Fusion of Multimers and Interface Mutations.

Nano letters·2015

Same journal

Wanted and unwanted modifications of mRNA, and their effect on gene expression and signaling.

The Journal of biological chemistry·2026

Same journal

TGF-β2 drives lipid droplet accumulation in chondrocytes through the TβRI/p-Smad3/Fabp5 axis.

The Journal of biological chemistry·2026

Same journal

Macrophage-specific targeting of histone demethylases with small-molecule inhibitors suppresses inflammatory response in vivo.

The Journal of biological chemistry·2026

Same journal

Substrate and target selectivity of 4'-fluoroadenosine against viral and host polymerases.

The Journal of biological chemistry·2026

Same journal

Correction: Characterization of Mast2 kinase defines structural features, regulation, and substrates.

The Journal of biological chemistry·2026

Same journal

Isotope-Edited ESEEM: A New Method for Probing Copper Binding Sites in Neurodegenerative Proteins.

The Journal of biological chemistry·2026

See all related articles

Search research articles

Related Experiment Video

Updated: May 30, 2026

Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA

Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA

Published on: November 14, 2017

Mining endonuclease cleavage determinants in genomic sequence data.

Mindy D Szeto¹, Sandrine J S Boissel, David Baker

¹Department of Biochemistry, Howard Hughes Medical Institute, University of Washington, Seattle, Washington 98195, USA.

The Journal of Biological Chemistry

|July 23, 2011

Summary

This summary is machine-generated.

Identifying new homing endonuclease variants is key for gene therapy. Researchers modified the I-AniI enzyme, creating novel variants with altered DNA targeting capabilities for improved gene editing tools.

More Related Videos

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

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

Published on: March 25, 2020

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

Published on: May 6, 2020

Related Experiment Videos

Last Updated: May 30, 2026

Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA

Simultaneous Mapping and Quantitation of Ribonucleotides in Human Mitochondrial DNA

Published on: November 14, 2017

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

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

Published on: March 25, 2020

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

Parallel High Throughput Single Molecule Kinetic Assay for Site-Specific DNA Cleavage

Published on: May 6, 2020

Area of Science:

Molecular Biology
Biotechnology
Genomics

Background:

Homing endonucleases are powerful tools for targeted gene therapy and gene correction.
Current limitations in identifying specific enzyme variants hinder their widespread application.
The LAGLIDADG homing endonuclease I-AniI serves as a scaffold for engineering new specificities.

Purpose of the Study:

To identify novel homing endonuclease variants with altered DNA target specificities.
To explore the potential of genomic data in redesigning endonuclease specificity.
To advance the development of precise gene editing tools for therapeutic applications.

Main Methods:

Homologues of the I-AniI endonuclease were identified using BLAST searches.
Genomic sequences flanking putative target sites were analyzed.
Amino acid substitutions conferring altered specificity were grafted onto the I-AniI scaffold.

Main Results:

Novel homing endonuclease variants were successfully generated.
Many engineered variants displayed new and unexpected DNA target specificities.
The study demonstrates the feasibility of exploiting genomic information for enzyme redesign.

Conclusions:

Genomic data can be effectively utilized to redesign homing endonuclease specificity.
Engineered endonucleases show promise for enhanced gene therapy and gene correction applications.
This approach facilitates the development of tailored DNA targeting tools.