Site-specific Integrase
Site-specific Integrase - Related Videos
08:21
Site-specific Bacterial Chromosome Engineering: ΦC31 Integrase Mediated Cassette Exchange (IMCE)
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Cited by 11 •
2012
A quick and efficient method to integrate foreign DNA of interest into pre-made acceptor strains, termed landing pad strains, is described. The method allows site-specific integration of a DNA cassette into the engineered landing pad locus of a given strain, through conjugation and expression of the ΦC31 integrase.
04:21
phiC31-Integrase-Mediated Site-Directed Transgene Integration: A Microinjection Technique for Site-Specific Transgene Integration into an Anopheles Vector
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2025
In this video, we demonstrate phiC31-mediated site-specific transgene integration in the Anopheles embryo via microinjection. This method helps introduce desired genes into a host system for research and industrial applications.
03:45
Purification of Viral Integrase Using Affinity Chromatography
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2026
Source: Lopez Jr., M. A., et al. Detection and Removal of Nuclease Contamination During Purification of Recombinant Prototype Foamy Virus Integrase. J. Vis. Exp. (2017)This video demonstrates the purification of polyhistidine-tagged viral integrase using nickel affinity chromatography. The protocol uses imidazole gradient elution to selectively isolate the integrase based on its affinity for nickel-charged resin.
02:21
Purification of Viral Integrase Using Heparin Affinity Chromatography
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2026
Source: Lopez Jr., M. et al. Detection and Removal of Nuclease Contamination During Purification of Recombinant Prototype Foamy Virus Integrase. J. Vis. Exp. (2017)This video demonstrates the procedure for purifying viral integrase from bacterial nuclease contamination using heparin Sepharose column chromatography followed by SDS-PAGE analysis to confirm protein purity.
07:50
Detection and Removal of Nuclease Contamination During Purification of Recombinant Prototype Foamy Virus Integrase
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Cited by 10 •
2017
Recombinant prototype foamy virus integrase protein is often contaminated with a bacterial nuclease during purification. This method identifies nuclease contamination and removes it from the final preparation of the enzyme.
FAQs
The integrase recognizes compatible attachment sequences, commonly described as att sites, rather than acting broadly across genomic DNA. This recognition brings the relevant DNA regions into the reaction and positions them for strand exchange. Because the enzyme depends on matching sequence signals, engineered DNA can be directed toward a defined genomic or construct location.
Strand exchange between recognized attachment sites changes the relationship between the DNA segments connected to those sites. Depending on how the sites and intervening DNA are arranged, the result can be insertion, excision, or inversion. This mechanism gives bioengineers several distinct ways to reorganize genetic material without relying on uncontrolled integration.
Random integration can place a transgene at variable genomic locations, whereas sequence-specific integrase activity is directed by compatible recognition sites. Defined targeting can reduce unwanted genomic disruption and make the resulting genetic change more reproducible. These features are especially important when engineered cells must maintain a consistent transgene arrangement or a controlled functional state.
A conceptual workflow begins by selecting compatible attachment sequences and positioning them around the DNA change to be controlled. The integrase is then used to catalyze strand exchange between those sites, producing the intended insertion, excision, or inversion. The resulting DNA arrangement can be examined to determine whether the planned genetic reorganization occurred.
Site-specific integrases support several bioengineering strategies, including stable transgene incorporation and modular genome construction. They also enable genetic memory circuits, in which a past cellular event can be represented through a lasting DNA arrangement, and controlled changes in cell function. These applications connect precise DNA remodeling with synthetic biology and biotechnology goals.
By linking DNA rearrangement to defined recognition sequences, integrase-based systems give researchers a way to encode controlled genetic changes in engineered cells. The resulting insertion, excision, or inversion can alter the organization of selected DNA and support changes in cell function. This programmability helps build cellular systems with more reproducible genetic behavior.