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Drosophila in vivo RNAi and Screening

RNA interference (RNAi) is a cellular mechanism of RNA-induced degradation using messenger RNA transcripts that can be used to silence gene expression and help identify and characterize gene function. RNAi has considerable applications in gene function detection and complements gene editing techniques in studying gene function. If your candidate gene is not suitable using knock-out technique, you can consider RNAi gene silencing strategy.

In our Drosophila in vivo RNAi service, gene function is systematically probed by large RNA hairpin fragments (300-400 bp) and short RNAs (shRNA, 80-250 bp) to disrupt the function of individual genes or on a large-scale. In addition, our RNAi Drosophila technique platform caters to in vivo large-scale or even genome-wide RNAi high-throughput screening to study fundamental biological processes, including cell signaling, environmental stress, the innate immune system, pain sensation, neuronal development, and human disease mechanism, etc.

CD BioSciences as your Drosophila research collaborator, provides you with reliable and rapid one-stop in vivo RNAi services to help your research projects advance. Microinjection is the simplest RNAi method. But this method is not suitable for all the tissues and genes, and the silencing results are usually unstable. We recommend using Gal4-UAS system to establish transgenic RNAi Drosophila strains to express dsRNA in a tissue-specific manner under Gal4 control. If your project requires, we also provide large-scale RNAi high-throughput in vivo screening services, including UAS-RNAi transgenic library construction, screening mechanism optimization, data analysis and off-target detection and analysis, etc.

Fig.1 RNAi Methods in Drosophila (Heigwer et al, 2018)

Feature Services
Workflow
Technical Characteristics

dsRNA microinjection

We offer Drosophila in vivo dsRNA injection services and qPCR validation, from embryos, larvae, pupae to adults. Customers can request to use your own dsRNA and lines. More conveniently, we can design and synthesize dsRNA directly for you, and inject Drosophila, as long as customers tell us the name of the gene. More details about our Drosophila microinjection service.

Establishing of transgenic UAS-RNAi Drosophila lines

Design of dsRNA sequence - We have a professional bioinformatic dsRNA construction platform to ensure sequence knockdown efficiency and minimize unintended sequence similarity. In addition, we construct 3 RNAi transgenic vectors against transcripts ruling out off-target effects. A variety of dsRNA lines (e.g. hsp70-, core promoter, UAS- Dcr-2, and selectable markers, etc.) are available to meet different experimental needs and prevent leaky activation or inefficiency.
Gal4 driver - The choice of Gal4 driver line is very critical for the efficiency of RNAi . The general recommendation is to select the Gal4 lines that are as strong as possible and start to be expressed before the phenotypic detection time point (e.g. Actin5C - or Tubulin-Gal4). Of course other Gal4 variants are also available (e.g. Gal80, HSP70-Gal4) to meet the more precise and flexible gene silencing requirements of our customers.
Validation and phenotypic analysis - Mature and perfect quality management process ensures the provision of genetically confirmed and functionally complete transgenic RNAi Drosophila strains to our customers. We will issue a results report for you.

Large-scale in vivo screening (optional)

Transgenic RNAi collection - We follow the standardized process to build transgenic RNAi libraries. We default to inserting all RNA vectors into the same genomic locus to overcome variability issues. Dicer enzyme co-expression genes are added in strains with lower absolute levels of RNAi.
Large-scale screening - Our professional team performs multiple pilot screens to determine the optimal screening phenotype and scoring rules before executing a large screen. Large screens are performed strictly according to quantitative and qualitative phenotypes, including microscopic observations, activity indicators, protein glycerol content, and other data. Finally, the quality of the obtained data is evaluated.
Bioinformatic date analysis - Gene ontology, comparison to existing high-throughput data, network and pathway analysis, etc.
Secondary screen(s) - Confirmation with different RNAi using the same analysis method; more detailed phenotypic analysis (antibody staining, real-time imaging, behavior, etc.); different Gal4 drivers to restrict cell types or stages.
Project delivery – The final report is mailed to you immediately after the experiment, containing the entire experimental procedure, certified results, screening strategy and bioinformatics analysis data, etc.

Almost all the genes can be silenced using RNAi.
In combination with the Gal4 drivers, gene knock-down can be controlled by time and space.
RNAi can mediate gene silencing at all developmental stages in Drosophila, including embryos, larvae, pupae, young and older adults.
Genome-wide RNAi screening is superior to traditional forward screening and can find hundreds of candidate positive genes.

Why CD BioSciences?

With many years of experience in Drosophila in vivo RNAi, we provide one-stop service from dsRNA design, vector construction, transgenic library construction, and bioinformatics analysis to meet the different needs of our global customers.
We provide reliable data and project reports for the entire experimental process. We guarantee reproducibility of all results.
With our own improved genome-wide RNAi screening platform, we can complete screening and data analysis of 10,000 - 20,000 RNAi lines in a short period of time.

CD BioSciences is an expert of gene silencing technology in fruit fly field. Our team of professionals provides 24 × 7 customer service to every client for saving his time and effort in biological research. There are no limitations of our service. If you need any further information or have any question, please feel free to contact us.

References

Kaya-Çopur A, et al. (2016). A Guide to Genome-Wide In Vivo RNAi Applications in Drosophila. Methods Mol Biol. 1478, 117–143.
Heigwer F, et al. (2018). RNA Interference (RNAi) Screening in Drosophila. Genetics. 208(3), 853–874.

For research use only. Not intended for any clinical use.

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