Combined Site-specific Recombinase System

Combined site-specific recombination systems refer to the use of multiple site-specific recombinase enzymes in a single experiment or application. By combining different recombinase systems, researchers can achieve more complex and precise genetic modifications that would be difficult to achieve with a single system. The combination of recombinases can also increase the efficiency and specificity of the genetic modifications, making them more useful for a wider range of applications.

What's Combined Site-specific Recombinase System

Site-specific recombinases are enzymes that catalyze the exchange of DNA fragments between two specific sites in a DNA molecule. The specificity of these enzymes is determined by the recognition sequence of the target sites, also known as recombination sites, in the DNA. When multiple site-specific recombinase systems are used, each recombinase recognizes and acts on a different set of recombination sites. This allows for the precise exchange and rearrangement of DNA fragments in a controlled manner.

Combining multiple site-specific recombinase systems offers a versatile tool for genetic engineering, as it provides greater control over the manipulation of DNA fragments. For example, the Cre/loxP system can be used to delete a specific gene or to insert a new gene into a desired locus, while the Flp/FRT system can be used to flip the orientation of a transgene. By combining these systems, researchers can perform multiple genetic modifications in a single experiment, increasing the efficiency and accuracy of their work.

Structural homology of tyrosine recombinase (Meinke, Gretchen et al. 2016)

Furthermore, using multiple site-specific recombinase systems in combination allows researchers to overcome some of the limitations of individual systems. For example, the Cre/lox system has the potential to induce genomic instability, while the Flp/FRT system requires the presence of a FRT site in the target DNA. By combining these systems, researchers can reduce the risk of genomic instability and increase the specificity of their manipulations.

What Can Combined Site-specific Recombinase System Do?

For example, the Flp/FRT system and the Cre/lox system are two commonly used site-specific recombinase systems. Flp/FRT recombinases can exchange a DNA fragment between two FRT sites, while Cre/lox recombinases can exchange a DNA fragment between two loxP sites. The use of different recombination sites in each system allows for the manipulation of different DNA regions without affecting the other. Multiple site-specific recombinase systems are widely used in various applications, such as gene expression control, genome engineering, and the production of genetically modified organisms.

Strategy for combining multiple site-specific recombinase system (Meinke, Gretchen et al. 2016)

Another important step in the precise shaping and remodeling of the genome is the combination of SSR with serine site-specific recombinase (S-SSR). Here, S-SSR is typically used in the first step to control the genomic insertion of DNA cassettes in the "safe harbor" region of the genome, while SSR is used in the second step to delete unwanted DNA. The combination of these two systems has been used to allow conversion of different overexpression alleles, combining the benefits of transgene targeting with modifiable transgene expression.

Why Combined Site-specific Recombinase System

As an additional optimization of traditional site-specific recombinase system, combined SSR system serve scientists as a more advantageous tool in genetic engineering, as well as other applications, offering advantages as following.

Increased Versatility: By combining different recombinase systems, researchers can design more complex genetic modifications that would be difficult to achieve with a single system. This increases the versatility of the recombinase systems for different applications.
Improved Efficiency: The use of multiple recombinases in combination can increase the efficiency of genetic modifications. For example, the Cre/loxP system has been shown to be more efficient in some applications when used in combination with the Flp/FRT system.
Improved Specificity: The combination of different recombinase systems can increase the specificity of genetic modifications. This is especially important in applications where it is important to avoid off-target effects.
Increased Flexibility: The combination of different recombinase systems can provide more options for controlling the timing and expression of genetic modifications, which can be useful in certain applications.

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Creative Biogene has years of experience in site-specific recombinase-based applications. We have established the advanced CreEdit^TM platform, which aims to support our global customers with high-quality, cost-effective and high-precision one-stop services. Our services are not limited in what we mentioned above, please feel free to contact us and get started with our first-class services.

Reference

Meinke, Gretchen et al. "Cre Recombinase and Other Tyrosine Recombinases." Chemical reviews vol. 116,20 (2016): 12785-12820.

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