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Chemical-inducible Site-specific Recombinase System
Inducible site-specific recombinase system is can be activated in response to specific environmental cues, such as the presence or absence of particular chemicals or changes in temperature. The activity of site-specific recombinase system can be controlled temporally and spatially by inducible recombinase system.
What's Chemical-inducible Recombinase System
A powerful addition to site-specific recombinase system is the availability of inducible recombinases, which allow scientists to manipulate DNA recombination within a specific period. The inducible site-specific recombinase system is a versatile tool with a wide range of applications in biotechnology, genetics, and other fields. Its ability to precisely manipulate DNA sequences in response to specific environmental cues makes it a valuable tool for advancing our understanding of gene function and regulation.
Chemical-inducible recombination systems (Tian X, et al. 2021)
Typical Chemical-inducible Site-specific Recombinase System
Tamoxifen-induced Cre-loxP System
Tamoxifen-induced Cre-loxP system is one of the most common inducible site-specific recombinase (site-specific recombinase) systems. The ubiquitous or tissue-specific promoter driven Cre is fused with estrogen receptor containing a mutated ligand binding domain (ER-LBD), as well as other nuclear receptor such as progesterone (PR), glucocorticoid (GR) or androgen (AR) receptors, which is normally bounded with heat shock protein 90 (HSP90) and the Cre-ER-HSP90 complex is prevented from entering the nucleus. Once the complex is bounded by synthetic analogs (tamoxifen, T, or 4-hydoxytamoxifen, 4-OHT), ER will be released from the HSP90 and enables the nuclear translation of Cre recombinase and allow the occurrence of recombinant reaction.
Tamoxifen-inducible Cre system (Hyeonhui Kim, et al., 2018)
Tetracycline/Doxycycline-induced Cre-loxP System
Tetracycline-induced Cre-loxP, as well as doxycycline (dox; a tetracycline derivative)-inducible system, are other well-known inducible site-specific recombinase systems. Three elements are involved in this system for controlling Cre recombinase, including reverse tetracycline-controlled transactivator (rtTA), tetracycline-controlled transactivator (tTA) and tetracycline responsive element (TRE). The system contains two modes: tet0on and tet-off, responsible for the activation and inactivation of the recombinase gene. In the tet-on mode, the rtTA binds to the dox to turn on the transcription of Cre gene. In contrast, the tTA binds to dox to turn off the transcription of Cre gene.
Other Chemical-inducible site-specific recombinase Systems
There are some other promising inducible-Cre system that are moving beyond the tamoxifen model and the dox model. The ER ligand binding domain is replaced by the progesterone receptor, which allows the induction with the progesterone analog RU-486. When the Cre is fused with a modified E. coli dihydrofolate reductase (DHFR) protein, the complex will be unstable and rapidly degraded in proteasomes in the absence of the inducer. Once treated with inducer, the common antibiotic trimethoprim (TMP), the protein will be stabilized and translocated to the nuclease and initiates the recombination reaction.
Advantages of Chemical-inducible SSR System
- Precision: One of the key advantages of it is its ability to precisely manipulate DNA sequences in response to specific environmental cues. This precision allows researchers to generate specific changes in the genome and study the effects of those changes on gene function and regulation.
- Versatility: It can be applied in a variety of cell types, including bacteria, yeast, plants, and mammals, making it a versatile tool for genetic manipulation across a wide range of organisms.
- Flexibility: It can be controlled by a variety of environmental cues, including changes in temperature, the presence or absence of chemicals, or exposure to light. This flexibility allows researchers to control when and where the recombination event takes place, making it a valuable tool for studying gene function and regulation in real-time.
- Cost-effective: The use of inducible site-specific recombinases is often more cost-effective than traditional methods of genetic manipulation, such as homologous recombination or transfection.
- Increased efficiency: It can be used to achieve high levels of efficiency in genetic manipulation, making it possible to study the function of multiple genes simultaneously.
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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
- Tian X, Zhou B. Strategies for site-specific recombination with high efficiency and precise spatiotemporal resolution. J Biol Chem. 2021;296:100509.
- Kim, Hyeonhui et al. Mouse Cre-LoxP system: general principles to determine tissue-specific roles of target genes. Laboratory animal research vol. 34,4 (2018): 147-159.