Technology
Online Inquiry
Split-Cre Recombinase System
Cre-loxP system is a very powerful tool for conditional genetic engineering, especially in mouse. There are hundreds of Cre mouse models available for performing tissue-specific KO in individuals. The split system is utilized by splitting Cre recombinase into two fragments (N-Cre and C-Cre) that are driven by separate promoters, which limited the expression of Cre in cells that express both Cre fragments. Furthermore, the activity of split-Cre system can be improved by the ligand inducing dimerization of complementary fragments fusing to N-Cre and C-Cre. The efficiency of DNA recombination is highly associated with the activity of Cre recombinase, which strictly depends on the affinity and stability of the dimer molecule fusing to N-Cre and C-Cre. Creative Biogene aims to optimizing the split-Cre system for improving the activity and controllability of genetic engineering.
What Is Split-Cre Recombinase System
Cre recombinase consists of 343 amino acids that form two domains, N-terminal domain and C-terminal domain. The N-terminal domain contains residues 20–129, which form five α-helical segments linked by short loops. The C-ternimal domain contains residues 132-341, which compose the active site of Cre recombinase. Based on its structure, Cre recombinase can be split into two complementation polypeptides at different break points, such as Asn59/Asn60, Leu104/Arg106 and Gly190/Gly191, with the ability of being reconstructed in vivo.
In the split-Cre system, two recombinase fragments have no detectable activity individually. However, the co-expression of two fragments lead to the reconstruction of an active recombinase. Many complementation systems have been utilized to reconstruct the N-Cre and C-Cre into active Cre recombinase, including intein-induced system, split-GPF-induced system, et al.
Increased Cre activity is detected while the N-Cre and C-Cre are reconstructed through GFP. As on of the most commonly used fluorescent proteins, the fusion of GFP and Cre is widely utilized for real-time labeling the expression of Cre recombinase.
Why Split-Cre?
The Cre-loxP system is commonly used in genetic engineering, but there are limitations to its accuracy. The specificity of Cre expression affects the technology's precision, as even small amounts of Cre expressed in non-targeted cells can confuse data interpretation or cause embryonic stem cell lethality. The split-Cre system has been developed to improve expression control by using N-Cre and C-Cre driven by different promoters, enhancing tissue-specific expression precision.
| Advantages of Split-Cre System | |
|---|---|
| Increased efficiency | The Split-Cre system allows for a more efficient generation of conditional knockout mice compared to traditional methods, as it reduces the number of mice that need to be bred and genotyped. |
| Increased specificity | The Split-Cre system allows for greater specificity in the activation of Cre recombinase, as the timing and location of recombination can be carefully controlled. |
| Reduced background recombination | The Split-Cre system reduces the chance of "background" recombination, where Cre recombinase is activated in unintended cells or tissues, as compared to traditional methods. |
| Increased temporal control | The Split-Cre system allows for greater temporal control over the activation of Cre recombinase, enabling researchers to more precisely control the timing of gene deletion. |
| Reduced phenotypic variability | The Split-Cre system results in reduced phenotypic variability, as the activation of Cre recombinase is more controlled, reducing the impact of genetic variations on the phenotype. |
Why Choose Us
One-stop Service
High-efficiency
Professional Team
High-quality
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
- Meinke, Gretchen et al. "Cre Recombinase and Other Tyrosine Recombinases." Chemical reviews vol. 116,20 (2016): 12785-12820.
- Wei, Xundong et al. "A Split-Cre system designed to detect simultaneous expression of two genes based on SpyTag/SpyCatcher conjugation and Split-GFP dimerization." The Journal of biological chemistry vol. 297,4 (2021): 101119.