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Drosophila miRNA Targets Discovery

In Drosophila, miRNA binds mainly to the 3'UTR of the target, and there are also binding sites in the coding region and 5'UTR of mRNA, with different sites and different efficiency of regulation. A single miRNA can regulate hundreds of targets, and identifying the targets is the key to reveal the function of miRNA. Drosophila is the model organism of choice for studying miRNAs, and advanced technology allows several miRNA targets to be found using multiple strategies.

CD BioSciences utilizes an advanced research team to provide complete miRNA target prediction and high-throughput screening services for our global customers, including 3'UTR, 5'UTR, gene promoters and coding sequences. As well as our Drosophila in vivo experimental platform, provides reliable results for functional target validation. We are committed to improving the success rate of our clients' experiments and saving time and effort.

Methods of miRNAs targets identification

Fig.1 Methods of miRNAs targets identification (Hausser et al, 2014)

General Strategies

  • Ago1 HITS-CLIP / PAR-CLIP / iCLIP
    Ago1 is a key component of the miRNA complex, and thus can be enriched for protein-mRNA complexes using immunoprecipitation. CD BioSciences combines high-throughput sequencing technology with miRNA isolated by crosslinking immunoprecipitation (HITS-CLIP or CLIP-Seq) to map mRNA-Ago1 binding sites in vivo, which can reach single-base resolution and is suitable for different tissues and developmental stages in Drosophila.
    Our microarray platform also offers PAR-CLIP (Photo Activated Ribonucleoside Enhanced CLIP) and iCLIP (Individual Nucleotide Resolution CLIP) services.
    PAR-CLIP is an alternative to HITS-CLIP that uses a photo-reactive 4-thiouridine (or analog) to more accurately map miRNA-mRNA interactions. iCLIP significantly improves pairing resolution to a single nucleotide but with reduced efficiency.

Technical Highlights

  • Captures protein-target cross-linked structures with increased stability and capture probability
  • Captures without disrupting biopsies
  • Reduced RNA contamination, low noise and higher resolution
  • Enables discovery of new or uncommon miRNA target sites
  • High throughput, allowing genome-wide screening
  • Smaller datasets compared to genomes for easier locus analysis and prediction
  • In silico Predictions
    CD BioSciences builds multiple prediction platforms and algorithms for miRNA standards to meet the needs and research objectives of different clients. We use conserved, target sequence contextual feature information or RNA-RNA hybridization and other features to improve prediction accuracy. Our in-house scalable miRNA interaction database expands target binding data of miRNAs to Ago1 complexes, enabling more extensive and accurate prediction of miRNA targets.
Workflow of Drosophila miRNA target in silico Predictions- CD BioSciences

Technical Highlights

  • Diversify screening criteria
  • Streamline validation efforts
  • Data cleaning and visual presentation
  • Customized solutions
  • In vivo Target Validation
    Prior to in vivo target validation, bioinformatic prediction must be performed to identify a list of candidate targets. Through our standardized genetic manipulation platform, recombinant reporter cell lines containing specific targets are typically constructed for miRNA transfection analysis experiments to determine the functional targets of miRNAs. We offer an integrated cell line validation service where you simply provide your list of target candidates.

Technical Highlights

  • Combine multiple analytical solutions such as biochemical immunology and imaging
  • High sensitivity and cellular resolution
  • Multiple reporter genes for analysis of different periods, tissues and other conditions
  • Customized reporting system one-stop solution service

Why CD BioSciences?

  • Rich experience in Drosophila miRNA research
  • Excellent and professional research team and technical platform
  • Flexible miRNA assay and bioprocess integration services
  • Reliable documented data, results and protocols
  • Fast turnaround, cost-effective and transparent pricing

CD BioSciences is an expert of miRNA research in fruit fly field. We provide integrated services from design, optimization, analysis to validation of miRNA targets. After the project is completed, we will mail the complete experiment report and raw data, and provide 24 × 7 technical support to save your effort and time. There are no limitations of our service. If you need any further information or have any question, please feel free to contact us.

References

  • Hausser J, et al. (2014). Identification and consequences of miRNA–target interactions — beyond repression of gene expression. Nat Rev Genet. 15, 599-612.
  • Riolo G, et al. (2021). miRNA Targets: From Prediction Tools to Experimental Validation. Methods Protoc. 4, 1.

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

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