Gal4 Gal80 System in Drosophila
The yeast galactose induction system, induced by the transcriptional activator Gal4 and its specific UAS binding site, has been well-appreciated a highly successful and versatile system for controlling transcriptional stimulation in Drosophila. However, different experimental purposes may require gene expressions or manipulations to be achieved in a more limited time window or with greater tissue specificity. Modification of Gal4 system in many ways refines the expression and produces more specific and precise patterns. Gal80 binding to Gal4 plays a role as an inhibitory factor to repress the transcription activity of Gal4. The expression of Gal4-dependent transgenes can be controlled accurately by the command of Gal80 variants.
CD BioSciences is an allied team of scientists and project management professionals dedicated to refining more useful fruit flies models supporting researches range from gene function to single pathway and development mechanism. For many years of exploration, 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 Gal4 Gal80 system?
Using Gal4-UAS system, with aid of multi-copies UAS at adjacent sites and specific Gal4 drivers, approaches ectopic activation of gene expression under researchers' control. Improvements to the Drosophila Gal4-UAS system have become more and more complex and personalized. It meets the various requirements that can be used to study the various stages of development, as well as the physiology and behavior of fly adults.
One of the modification strategies is transferring yeast Gal80 gene, which binds to Gal4 activation domain and blocks its interaction with UAS-regulated transcriptional machinery. Distinct Gal80 Gal4 systems attain the precise expression patterns of Gal4-dependent transgenes propelling the Temporal and Regional Gene Expression Targeting (TARGET) technology. Gal80 crosses are much more convenient and time-saving than classical breeding approaches using null mutations screening. The most commonly regulating factor is temperature. Temperature-sensitive alleles of Gal4 itself have been generated and tested in Drosophila, however the ease of combining Gal80TS with already established Gal4 drivers cause the TARGET system to be more flexible used. Gal80TS is a temperature-mediated variant. Alternatively, combining a heat shock promoter also approaches Gal4-dependent gene expression in temperature activation. At permissive temperatures (18-20℃), Gal80TS binds to Gal4, whereas at restrictive temperatures (29-30℃), it no longer combines and Gal4-mediated transcriptional activation is recovered.
It is worth noting that, the Gal80 Gal4 system not only controls the expression of the target gene instantaneously and accurately, but also achieves the flexible regulation of the gene expression level under effectively control of the intensity of stimulus (temperature and light, etc.). It is helpful to explore the different functions of some genes with concentration effects, or the genes responding different function in different cell types.
Modification Gal80 Gal4 system is specific and personalized, our services are not merely temperature activation system. We try our best to provide the optimal refine strategies in accordance with the requirements of our customers.
Workflow of constructing and optimization of Gal4 Gal80 system
- MODEL STRATEGY
- EGG INJECTIONS
- CANDIDATE SCREENING
We collaborate with you to understand your final research expectations. This allows us to make a comprehensive plan, detailing inserting sites, various Gal80 modification schemes, optimizing Drosophila codon sequences, assembling optimal vector, Gal4 lines selection to maximize the chance of success. We provide you with the most convenient and effective strategy for your research purpose.
Depending on the project approach, we offer optimal vector tool and Gal80 variants (optional) generation through our suppliers. Besides our Gal4 and UAS lines stock, we deploy a variety of gene assembly approaches to get the exact sequences and plasmids (usually pUAST) needed for appropriate Gal80 Gal4 system. Moreover, we can take advantage of Pelement transposition and enhancer trap vectors to integrate Gal4 gene randomly into the fly genome to design your own Gal4 line resources.
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.
Once we confirm the ends of the gene insertion have proceed as planned, strains homozygous/heterozygote are crossed. Depending on different project, we have G0 and F1 screening by "visible" marker or sequence validation.
G0 or/and F1 are positive screened and confirmed by full sequencing of the entire transgene sequence due to client's opinion. As standard, we validate at the genomic level with advanced sequencing and we provide a functional readout that verifies conditional gene KO efficiency.
During the fruit flies engineering process, our team performs the desired edit(s) defined in the statement of work. 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.
- Study the biological functions of certain genes/proteins at the tissue-specific/cellular level
- Study brain functions such as learning and memory
- High-throughput drug screening and development
- Verify targets drug through Drosophila model
- Preclinical safety and toxicity studies
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 vector construction and library screening.
CD BioSciences as a research-driven biotechnology company continuously improving upon the standards we set for ourselves and our industry. We offer one-stop Gal80 Gal4 system construction and modification strategy for gene function and mechanism researches in Drosophila. Because each project is different, if you could not find what you really want in the website, please feel free to contact us. Our project management team will return to you as soon as possible.
For research use only. Not intended for any clinical use.