Recombinase Technology
Recombinase Technology - Related Videos
07:06
Principles of Site-Specific Recombinase (SSR) Technology
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Cited by 14 •
2008
The advent of site-specific recombinase (SSR) technology and the Cre/lox system has led to numerous advances in molecular biology, and has proven itself as a valuable tool for assessing gene function in transgenic animals. This interview discusses the mechanism of site specific recombination by Cyclization recombinase (Cre) and how the use of this enzyme has led to the development of conditional mutagenesis, which has significant advantages over traditional knock out strategies.
12:02
Molecular Evolution of the Tre Recombinase
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Cited by 4 •
2008
Here we report the generation of Tre recombinase through directed, molecular evolution. Tre recombinase recognizes a pre-defined target sequence within the LTR sequences of the HIV-1 provirus, resulting in the excision and eradication of the provirus from infected human cells. While still in its infancy, directed molecular evolution will allow the creation of custom enzymes that will serve as tools of molecular surgery and molecular medicine.
10:20
Interview: HIV-1 Proviral DNA Excision Using an Evolved Recombinase
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Cited by 2 •
2008
Current HIV-1 strategies act to suppress the viral life cycle but do not effectively eradicate infection. Here, we demonstrate that an engineered recombinase can efficiently excise integrated HIV-1 proviral DNA from the genome of infected cells.
03:12
CreER-LoxP System-Based Target Gene Inactivation: A Tamoxifen-Inducible Cre Recombinase System for Target Gene Knockout in a Mouse Model
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2025
This video describes a procedure for target gene knockout using a tamoxifen-inducible Cre-recombinase system following intraperitoneal injection of tamoxifen in a mouse model.
15:13
Structure-function Studies in Mouse Embryonic Stem Cells Using Recombinase-mediated Cassette Exchange
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Cited by 6 •
2017
Proteins often contain multiple domains that can exert different cellular functions. Gene knock-outs (KO) do not consider this functional diversity. Here, we report a recombination-mediated cassette exchange (RMCE)-based structure-function approach in KO embryonic stem cells that allows for the molecular dissection of various functional domains or variants of a protein.
FAQs
The arrangement of recognition sites determines the DNA outcome. When matching sites flank a segment in a configuration that permits removal, the intervening sequence can be excised. Other orientations can reverse the segment rather than remove it, while different site arrangements can support insertion. Thus, site placement is a design variable, not merely a targeting detail.
Cre and FLP provide alternative site-specific recombinase systems, but each recognizes its corresponding DNA element: loxP for Cre and FRT for FLP. A construct must therefore pair the enzyme with the matching sites rather than treating recognition sequences as interchangeable. This modularity lets researchers select a compatible system for a genetic design and maintain defined control over recombination.
Conditional knockout depends on restricting where or when recombination occurs. A target gene can be arranged with recognition sites, while recombinase activity is limited to selected cells or contexts. Recombination then changes the target sequence only in that controlled setting, allowing comparisons between affected and unaffected cells. This is useful for examining gene function without treating every cell identically.
Orientation is important when the experimental goal is sequence inversion rather than deletion. Recognition sites arranged in opposite orientations can redirect the intervening DNA, whereas a different arrangement can enable excision or insertion. The expected product therefore depends on both the selected recombinase system and the geometry of its sites. Mapping that geometry helps interpret the resulting DNA change.
A practical design begins by choosing a matching recombinase and recognition-site pair, then placing those sites around or within the DNA sequence whose behavior must be controlled. Researchers next arrange the sites to produce the intended outcome, such as excision or inversion, and apply the system in selected cells or model organisms. The resulting DNA change can then be evaluated in its biological context.
Recombinase technology is useful when a study requires gene function to be examined in a defined cellular or developmental context. Its controlled rearrangements support conditional gene knockout, transgene activation, and lineage tracing. These applications let investigators connect a genetic change with particular tissues or stages, strengthening analyses of development, disease mechanisms, and tissue-specific biology.
Lineage tracing uses controlled recombination to connect a cell population with a defined genetic rearrangement. When recombinase activity is limited to selected cells, the resulting DNA configuration identifies which cells experienced the event. Researchers can then relate that population to developmental or tissue-specific processes, making the approach valuable for studying how biological roles are distributed across cells.