Site Specific Recombination
Site Specific Recombination - Related Videos
02:53
Conservative Site-specific Recombination and Phase Variation
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2020
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...
14:23
Recombineering Homologous Recombination Constructs in Drosophila
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Cited by 9 •
2013
Homologous recombination techniques greatly advance Drosophila genetics by enabling the creation of molecularly precise mutations. The recent adoption of recombineering allows one to manipulate large pieces of DNA and transform them into Drosophila6. The methods presented here combine these techniques to rapidly generate large homologous recombination vectors.
08:56
Recombineering and Gene Targeting
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2023
One of the most widely used tools in modern biology is molecular cloning with restriction enzymes, which create compatible ends between DNA fragments that allow them to be joined together. However, this technique has certain restrictions that limit its applicability for large or complex DNA construct generation. A newer technique that addresses some of these shortcomings is recombineering, which modifies DNA using homologous recombination (HR), the exchange between different DNA molecules based...
03:11
Recombinant Adeno Associated Virus Based Gene Delivery: A Technique for Transgene Delivery and Cardiac-Specific Expression in Rodent Model
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2025
In this video, we demonstrate recombinant adeno associated virus-based gene delivery in the mouse pup. This method helps in studying gene expression in the targeted cells.
11:25
Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins
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Cited by 1 •
2017
Biochemical and structural analyses of glycosylated proteins require relatively large amounts of homogeneous samples. Here, we present an efficient chemical method for site-specific glycosylation of recombinant proteins purified from bacteria by targeting reactive Cys thiols.
FAQs
The decisive variables are the orientation and location of the recognition sites. When sites are arranged in different configurations, the same recombination machinery can generate integration, excision, inversion, or replacement. Site design therefore does more than identify where recombination occurs: it determines how the DNA segment will be reorganized and what genetic change becomes possible.
The recombinase provides molecular selectivity by binding the appropriate short recognition sequences. It brings the corresponding DNA molecules or regions into alignment and catalyzes strand exchange between them. This coordinated action restricts the genetic rearrangement to defined sites, allowing researchers to make targeted changes rather than relying on an unspecified location within the genome.
A compatible recombinase and recognition-site pair establishes which DNA regions can participate in the reaction. Systems such as Cre–lox and Flp–FRT use this pairing to support controlled genetic modification. The pairing is especially useful when researchers need a defined genetic change, because the selected sites provide the positional information that guides the rearrangement.
Researchers assess where the recognition sites are located and how they are oriented relative to the DNA segment of interest. These features provide a basis for anticipating the rearrangement produced after strand exchange. Planning the site arrangement in advance helps align the recombination design with the intended genetic outcome, such as removing, reversing, inserting, or replacing material.
An experiment first establishes the relevant recognition sequences around or within the DNA region to be modified. The matching recombinase system is then used to bind those sites, align the DNA regions or molecules, and catalyze strand exchange. Researchers interpret the resulting integration, excision, inversion, or replacement as the designed genetic modification.
In conditional knockout models, recombination provides a way to control gene modification in selected cells or under selected conditions. Researchers can use a compatible recognition-site system to target the relevant genetic material and then examine the resulting change in gene function. This approach helps separate a gene’s effects from broader, unrestricted modification.
Lineage tracing uses controlled genetic modification to mark or alter genetic material in selected cells, allowing researchers to follow the consequences in those cellular lineages. Site-specific systems contribute the positional precision needed for this strategy. The resulting genetic record can help investigators study how selected cells and their descendants contribute to biological processes.
Site-specific recombination also supports transgene integration and regulation of gene expression in selected cells or conditions. These uses let researchers introduce or control genetic material with greater precision than an unrestricted modification would provide. Together with knockout and lineage-tracing applications, they create tools for investigating gene function and engineering targeted genetic changes.