Split Gal4 System in Drosophila
Gal4-UAS system has been used to direct the expression of targeted gene in Drosophila. Gal4 shows to possess distinct protein domains for binding to DNA (DBD) and for activating transcription (AD). When Gal4 identifies and combines to UAS, the UAS-targeted gene will be expressed. While, Split Gal4 system is a derivative of Gal4-UAS system. In simple terms, the system expresses the two important domains (DBD and AD) separately, and only when the two parts are combined can reconstitute Gal4 activity activating the transcriptional function.
CD BioSciences is a leading technology-driven custom service provider majoring in many fields related fruit fly. With past decade of efforts, we have established a complete platform for refining and manufacturing customized Gal4-UAS system in Drosophila, beyond the scope of Gal80 variants, GeneSwitch, and Split Gal4 system. We offer the best quality solutions to facilitate the development of your researches and projects.
What is Split Gal4 system?
Another spatial-refined system is Split GAL4 system. As the name, this system remains only 2 parts of Gal4 primary domains driven by different enhancers, the DNA binding domain and transcriptional activation domain. Only if both "hemi-divers" are expressed and fused correctly in the same cell, the transcription of UAS-gene can be activated. Through the improvement of the connection part recognition and hemi-drivers' expression methods, the selective expression for different purposes can be achieved. In general, the DBD and AD of Gal4 are fused independently to heterodimerizing synthetic leucine zippers via a polyglycine spacer. But these years, several new combinations have been raised. This kind of refine reduces an extent of leak and allows cells manipulation. Obviously, Split Gal4 lines can refine expression patterns to very small numbers of cells, allowing for example, the manipulation of small groups of neurons in behavioral studies. This combinatorial system realizes a high degree of spatial refinement allowing multiple refine transcriptional strategies.
Workflow of constructing and optimization of Split Gal4 system
- MODEL STRATEGY
- HEMI-DRIVER CONSTRUCTS
- EGG INJECTIONS
- CANDIDATE SCREENING
We provide you with the most convenient and effective strategy for your research purpose. This step allows us to make a comprehensive plan, detailing various modification schemes, optimizing Drosophila codon sequences, assembling optimal vector, selection of lines with insertion site to maximize the chance of success. Moreover, we provide one-stop specific-tissue driver screening (neuro, intestinal cells, etc.) with gene trap or enhancer trap.
Depending on the project approach, we offer optimal hemi-driver lines generation. Besides our hemi-drivers and enhancers library, we provide specific-tissue driver screening with evaluation of the expression patterns to improve hemi-drivers constructs for appropriate Split Gal4 system. We generally use ϕC31 site-specific integrase into the attP landing site and Cas9 nuclease introducing highly specific genes.
Up to 300-embryos are microinjected. Our injection operators have infallible skills and experience. They can obtain precision delivery of transgenesis reagents to Drosophila embryos. We perform a sufficient number of injections that promising >100 crossed G0. And raise the survived larvae to adulthood.
In generally, an exogenous marker or reporter helps identify candidates. But there are no limits in screening pathways. For our genomic integrant, we look at each transgenesis (including effector and transactivator transformants) by PCR.
A series of genetic crosses to remove the integrase source, to establish a homozygous stock and to construct stable Split Gal4 lines that carry a combination of the two hemi-drivers. As standard, we validate at the genomic level with advanced sequencing and we provide a functional readout that verifies system efficiency and flies' vitality.
We provide regular updates and in-time messages if any issue or complication happen during the manufacturing process.
We ship final transformants in either larvae or adults. Technologically, balancing is necessary if the mutation is homozygous lethal or sterile. We send you F1 homozygous or balanced stable transformants flies. The final transformants vials are maintained in the facility for two weeks as backup.
- Identification of protein-protein interaction domains
- Gene function and signal regulated conduction
- Characterizing cell groups both in physiology and structure, for example, neurons of interest.
- Functional screens
Why CD BioSciences?
- Record-proven expertise and experience in Gal4-UAS system and Drosophila.
- Consistent results and superior efficiency.
- Fast turnaround services.
- Best after-sale service in construction and library screening.
CD BioSciences is devoted to providing highly customized all-round solutions with the best quality to support our global clients' projects. We offer one-stop Split Gal4 system constructs and modification strategy for gene function and mechanism researches in Drosophila. For more information, don not hesitate to contact us, you can quickly attain your high-quality Split Gal4 Drosophila lines and study gene expression in many different tissue types!
For research use only. Not intended for any clinical use.