Conditional Knockout (KO) Service

Conditional knockout (cKO) is a technique used in molecular biology to genetically modify an organism, typically a mouse, to study the function of a specific gene. The cKO models have provided valuable insights into the functions of genes in various biological processes, including development, disease, and behavior. Creative Biogene offer conditional KO services in multiple organisms for research use.

Introduction into Conditional Knockout (cKO)

Conditional knockout (cKO) is a genetic technique used to study the function of a specific gene in a particular tissue or at a particular developmental stage in a living organism. In a traditional knockout experiment, the entire gene is deleted in all cells of the organism, which can result in embryonic lethality or pleiotropic effects that make it difficult to discern the gene's specific role in a particular tissue or developmental stage.

In contrast, a conditional knockout experiment allows for more precise control over when and where a gene is inactivated. This is accomplished by introducing a genetic modification that allows the gene to be deleted in a specific tissue or at a specific time. This can be done using a variety of genetic techniques, including the Cre-loxP system or the tamoxifen-inducible Cre-ER system.

Tamoxifen-inducible Cre-loxP mouse knockout model (Smith, Lorna I F et al., 2020)

The ability to selectively delete a gene in a particular tissue or at a particular time is valuable for investigating the gene's function in specific contexts. For example, a researcher might be interested in studying the role of a particular gene in the development of the heart, but deleting the gene in all cells of the organism might lead to embryonic lethality before the heart has a chance to develop. By using a cKO approach, the gene can be deleted specifically in the heart tissue, allowing the researcher to study the gene's role in heart development without affecting other tissues or embryonic development.

Technologies for cKO

The most common and efficiency technology for cKO is the site-specific recombinase system, especially the Cre/loxP system. Creative Biogene has developed various of technologies based on site-specific recombinase system for meeting each specific scientific question. Followings are introduction of commonly used site-specific recombinase systems:

• Cre/loxP system

This system uses the Cre recombinase enzyme and loxP sites to induce DNA recombination. It is widely used and well-established in mouse genetics. The main advantage of this system is its versatility and specificity, as the Cre enzyme can recognize loxP sites and efficiently excise or invert DNA sequences flanked by them. However, the size of the loxP sites may limit its application in certain organisms or genomic regions.

• Flp/FRT system

This system uses the Flp recombinase enzyme and FRT sites for DNA recombination. It is similar to the Cre/loxP system in terms of mechanism and efficiency, but has smaller site sizes which allow for more flexibility in genome engineering. It is commonly used in Drosophila genetics.

• PhiC31/attP-attB system

This system uses the PhiC31 integrase enzyme and attP and attB sites for DNA integration. It allows for precise integration of DNA sequences into the genome, and has been used in both Drosophila and mammalian systems. One disadvantage is that the attP and attB sites are not naturally present in mammalian genomes.

• Dre/rox system

This system uses the Dre recombinase enzyme and rox sites for DNA recombination. It has been used successfully in Drosophila and has the advantage of smaller site sizes than the Cre/loxP system.

Our Service

Conditional KO models are powerful tools for gene functional study. Creative Biogene offers customized cKO service for our global customers with the well-developed T-SSR-based technologies.

Why Choose Us

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

1. Smith, Lorna I F et al. "Generating Beta-Cell-Specific Transgenic Mice Using the Cre-Lox System." Methods in molecular biology (Clifton, N.J.) vol. 2128 (2020): 181-205.

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