Recombinase Engineering
Recombinase Engineering - 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.
09:36
Genetic Engineering of Model Organisms
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2023
Transgenesis, or the use of genetic engineering to alter gene expression, is widely used in the field of developmental biology. Scientists use a number of approaches to alter the function of genes to understand their roles in developmental processes. This includes replacement of a gene with a nonfunctional copy, or adding a visualizable tag to a gene that allows the resultant fusion protein to be tracked throughout development. In this video, the viewers will learn about the principles behind...
FAQs
Engineering can modify which DNA sequences a recombinase recognizes, allowing researchers to redirect activity toward a selected genetic site. This sequence specificity determines where rearrangements occur, while optimization of activity and stability helps the enzyme function effectively in the intended system. Together, these changes support more controlled manipulation of genetic information than an unoptimized enzyme may provide.
The arrangement of the recognized DNA sites determines the type of genetic rearrangement produced. Integration adds a genetic element at a selected location, whereas excision removes one. Inversion changes the orientation of a DNA segment, and translocation moves genetic material between locations. Engineering reaction direction can therefore determine whether a system inserts, removes, flips, or relocates a sequence.
Activity affects how effectively the enzyme catalyzes strand exchange, while stability influences whether it remains functional in the target biological system. Adjusting these properties helps match recombinase performance to a particular experimental context. Optimization is important because a useful design must not only recognize the intended sequence but also produce the desired rearrangement reliably.
A recombinase can be incorporated into a synthetic gene circuit so that recognition of defined DNA sites changes the arrangement of genetic elements. Depending on the engineered reaction, this rearrangement can alter the state of the circuit through integration, excision, or inversion. Such switches let researchers build cell models in which selected genetic states can be controlled and studied.
A typical workflow begins by identifying the genetic site and rearrangement required for the experiment. Researchers then design or optimize the recombinase for the needed sequence specificity, reaction direction, activity, or stability, and introduce the system into the target model. The resulting genetic rearrangement can then support analysis of gene function, lineage behavior, or circuit state.
The approach is useful when researchers need controlled genetic changes at selected sites rather than unrestricted modification of genetic information. Applications include genome editing, transgene integration, synthetic gene circuits, and lineage tracing. It also supports programmable cell models for investigating gene function and contributes to biotechnology and therapeutic research by making genetic systems more tunable.