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

Conservative Site-specific Recombination and Phase Variation

Conservative Site-specific Recombination and Phase Variation

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...

CRISPR

CRISPR

Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...

RNA Splicing

RNA Splicing

Splicing is the process by which eukaryotic RNA is edited before its translation into protein. The RNA strand transcribed from eukaryotic DNA is called the primary transcript. The primary transcripts that become mRNAs are called precursor messenger RNAs (pre-mRNAs). Eukaryotic pre-mRNA contains alternating sequences of exons and introns. Exons are nucleotide sequences that code for proteins, whereas introns are the non-coding regions. In RNA splicing, introns are removed and exons are bonded...

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...

You might also read

Related Articles

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

Sort by

Same author

A retargeted recombinase for precise insertion of large DNA.

Nature biotechnology·2026

Same author

Development of Cathepsin B‑Activatable Cell-Penetrating Peptides for Tumor Targeting.

ACS pharmacology & translational science·2026

Same author

Glycosaminoglycan-functionalized hydrogels for sustained delivery of tissue inhibitor of metalloproteinase-3 mediating matrix metalloprotease inhibition and extracellular matrix stabilization.

Bioactive materials·2026

Same author

<i>In Silico</i> Analysis of Serum Albumin Binding by Bone-Regenerative Hyaluronan-Based Molecules.

Pharmaceutics·2025

Same author

Heparan sulfate fine-tuned interleukin-1 (IL-1) signaling inhibits insulin secretion of grafted pancreatic islets.

Science advances·2025

Same author

A base editing platform for the correction of cancer driver mutations unmasks conserved p53 transcription programs.

Genome biology·2025

Search research articles

Related Experiment Video

Updated: Jun 24, 2025

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

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

Published on: March 29, 2019

Engineering spacer specificity of the Cre/loxP system.

Jenna Hoersten¹, Gloria Ruiz-Gómez², Maciej Paszkowski-Rogacz¹

¹Medical Faculty and University Hospital Carl Gustav Carus, UCC Section Medical Systems Biology, TU Dresden, 01307 Dresden, Germany.

Nucleic Acids Research

|June 13, 2024

Summary

Researchers explored the Cre/loxP system

More Related Videos

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

Principles of Site-Specific Recombinase SSR Technology

Principles of Site-Specific Recombinase SSR Technology

Published on: May 29, 2008

Related Experiment Videos

Last Updated: Jun 24, 2025

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

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

Published on: March 29, 2019

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Subcloning Plus Insertion SPI - A Novel Recombineering Method for the Rapid Construction of Gene Targeting Vectors

Published on: January 8, 2015

Principles of Site-Specific Recombinase SSR Technology

Principles of Site-Specific Recombinase SSR Technology

Published on: May 29, 2008

Area of Science:

Molecular Biology
Genetics
Biotechnology

Background:

The Cre/loxP system is a widely used tool for genetic manipulation.
Enhancing Cre/loxP specificity is crucial for precise genome engineering.
Mechanisms of Cre's substrate discrimination are not fully understood.

Purpose of the Study:

To investigate the impact of spacer sequence variations on Cre/loxP recombination efficiency.
To explore the potential for reprogramming Cre recombinase specificity.
To understand the molecular basis of Cre-DNA interactions.

Main Methods:

Screening of 6000 unique loxP-like spacer sequences.
Directed evolution of Cre recombinase.
Molecular modeling and dynamics simulations.

Main Results:

Approximately 84% of tested spacer sequences supported efficient recombination, indicating significant plasticity.
Specific spacer sequences were found to negatively impact recombination efficiency.
Directed evolution generated Cre variants with altered and enhanced spacer specificity.

Conclusions:

Spacer sequence in loxP sites significantly influences Cre recombination efficiency.
Cre recombinase activity and specificity can be reprogrammed through directed evolution.
This work provides a foundation for developing designer recombinases with fine-tuned specificity for genome engineering.