Related Experiment Video
Updated: Jun 20, 2025
07:06
Principles of Site-Specific Recombinase SSR Technology
Published on: May 29, 2008
20.9K
Dynamics in Cre-loxP site-specific recombination.
Mark P Foster1, Matthew J Benedek1, Tyler D Billings1
1Department of Chemistry and Biochemistry, The Ohio State University, Columbus, OH, USA.
Current Opinion in Structural Biology
|July 19, 2024
Summary
Cre recombinase precisely manipulates DNA by recognizing loxP sites. Understanding the dynamic properties of Cre and DNA is key to controlling its activity for advanced genetic applications.
Area of Science:
- Molecular Biology
- Biochemistry
- Genetics
Background:
- Cre recombinase is a widely used enzyme for site-specific DNA recombination.
- Its applications are limited by incomplete understanding of target recognition and activity control.
- Cre-loxP recombination is a powerful tool for genetic engineering.
Purpose of the Study:
- To elucidate the mechanisms of Cre-loxP target recognition and DNA cleavage.
- To investigate the role of dynamic properties in Cre recombinase activity.
- To identify strategies for controlling Cre activity for broader applications.
Main Methods:
- High-resolution structural and biophysical methods were employed.
- Analysis focused on the dynamic interplay between Cre enzyme and loxP DNA targets.
- Investigated sequence recognition and DNA cleavage steps.
Main Results:
- Dynamic properties of both Cre and its DNA target are critical determinants of recombination.
- Understanding these dynamics is essential for both sequence specificity and cleavage efficiency.
- Emerging data highlights the importance of enzyme-DNA dynamics.
Conclusions:
- Improved understanding of Cre-loxP dynamics will enable redirection of Cre to new sequences.
- This knowledge will facilitate better control over Cre DNA cleavage activity in vitro and in vivo.
- Advances in understanding enzyme-DNA dynamics promise enhanced genetic engineering tools.
Related Concept Videos
02:53Conservative 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...
The recognition sites for Cre recombinase called LoxP...
6.0K
02:31Homologous Recombination
4.5K
4.5K
01:30Crossing Over
4.3K
Crossing over is the exchange of genetic information between homologous chromosomes during prophase I of meiosis I. Genetic recombination gives rise to allelic diversity in the newly formed daughter cells. In humans, crossing over produces genetically distinct haploid egg and sperm cells that undergo fertilization to produce unique offspring. Before cell division starts, the germ cell’s chromosome(s) undergo duplication in the S phase of the cell cycle. As the cells enter prophase I,...
4.3K
02:32Exon Recombination
3.6K
The evolution of new genes is critical for speciation. Exon recombination, also known as exon shuffling or domain shuffling, is an important means of new gene formation. It is observed across vertebrates, invertebrates, and in some plants such as potatoes and sunflowers. During exon recombination, exons from the same or different genes recombine and produce new exon-intron combinations, which might evolve into new genes.
Exon shuffling follows “splice frame rules.” Each exon...
Exon shuffling follows “splice frame rules.” Each exon...
3.6K
01:02Long-patch Base Excision Repair
7.0K
Since the discovery of the two BER pathways, there has been a debate about how a cell chooses one pathway over the other and the factors determining this selection. Numerous in vitro experiments have pointed out multiple determinants for the sub-pathway selection. These are:
7.0K
02:08Gene Conversion
9.7K
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...
9.7K