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Combination of SSR and Other Tools
The combination of SSR with other techniques, such as RNA interference (RNAi), CRISPR/Cas, TALENs, or Zinc finger nucleases provides a means of creating precise and controlled genetic modifications in a variety of organisms. By using SSR to control the expression or activity of these techniques, researchers can achieve highly specific, efficient, and controlled genetic modifications in a variety of tissues or cell types, making the SSR system a powerful tool for basic research, biotechnology, and therapeutic applications.
Introduction into This Technology
Combining site-specific recombinase system together with other genetic engineering tools is a powerful technology that enables researchers to achieve greater precision and control over gene expression. Genetic engineering tools, such as Cre/loxP, Zinc finger nucleases, TALENs, or RNAi, are used to target specific genes and to manipulate their activity. By combining these technologies, researchers can achieve greater specificity, increased efficiency, and improved temporal control over gene expression.
Strategies of Combining SSR System and Other Tools
SSR and CRISPR/Cas
Combining Cre/loxp and CRISPR/Cas systems can provide a more versatile and efficient approach to making precise genetic modifications. The basic strategy involves using the CRISPR/Cas system to introduce loxP site into a specific locus, followed by the expression of Cre recombinase to excise the targeted DNA sequence.
SSR with Zinc finger nucleases or TALEN
Combining Cre/loxp with Zinc finger nucleases or TALEN is a strategy that allows for precise and controlled genetic modifications. The basic strategy involves using Zinc finger nucleases to introduce a double-stranded break (DSB) at a specific locus, followed by the expression of Cre recombinase to excise the targeted DNA sequence.
SSR and RNAi
The combination of Cre/loxP and RNAi can be used to achieve precise and controlled gene silencing in specific tissues or cells. The basic idea is to use Cre/loxP to drive the expression of an RNAi construct, such as a short hairpin RNA (shRNA) or a microRNA (miRNA), in a specific tissue or cell type. The RNAi construct is designed to target and silence a specific gene of interest, thereby allowing researchers to study the function of that gene in a highly controlled manner.
Why Do We Need to Combine SSR System with Other Tools
SSR system has been playing a very important role in genetic engineering during last few decades. The first step of applying this technology is introducing the target sites into genome, in which the homologous recombination (HR) technology was commonly used. However, the HR is relatively inefficiency in most cells. The application of nuclease significantly improved the production of conditional knockout alleles in several species. The combination of SSR with other nucleases provides several advantages, including:
- Increased specificity: The combination of SSR with other nucleases, such as CRISPR/Cas, TALENs, or Zinc finger nucleases, allows for the creation of highly specific and targeted genomic modifications. The ability of SSR to restrict the activity of these nucleases to specific sequences in the genome enhances their specificity, allowing for the creation of precise and controlled genetic modifications.
- Increased efficiency: The combination of SSR with other nucleases can increase the efficiency of genomic modifications, as the nuclease can be directed to the precise location in the genome where the modification is desired.
- Improved scalability: The combination of SSR with other nucleases can make it easier to scale up the production of genetically modified organisms, as the use of a combination of systems can simplify the process of creating genetic modifications and increase the efficiency of the process.
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Creative Biogene has years of experience in site-specific recombinase-based applications. We have established the advanced CreEditTM 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.
References
- Horii, Takuro et al. Efficient generation of conditional knockout mice via sequential introduction of lox sites. Scientific reports vol. 7,1 7891. 11 Aug. 2017
- Li, H., Yang, Y., Hong, W. et al. Applications of genome editing technology in the targeted therapy of human diseases: mechanisms, advances and prospects. Sig Transduct Target Ther 5, 1, 2020.