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Library-based Functional miRNA Screening is an innovative, high-throughput phenotype-driven service designed to systematically identify functional miRNAs that regulate specific biological processes. Unlike traditional screening methods based solely on differential expression, Creative Biogene's platform directly observes the functional impact on cellular phenotypes caused by over-expression or inhibition of miRNAs, offering clients biologically meaningful regulatory factors.
Creative Biogene's service is built on a robust technical platform composed of several core components. First, our high-quality miRNA intervention libraries include two main options:
miRNA Mimics library
Chemically synthesized double-stranded RNA molecules that precisely mimic mature miRNAs for over-expression studies, covering over 900 miRNAs across human, mouse, and other species.
miRNA Inhibitors library
Chemically modified single-stranded RNAs that specifically bind and inhibit endogenous miRNA function for loss-of-function screening. Each library undergoes stringent quality control to ensure high purity, minimal off-target effects, and consistent batch-to-batch stability, and includes essential negative and positive controls. Clients can visit the broader miRNA Service section for additional context.
In the selection of cell models, we recommend cell lines most suitable for the target phenotype based on the client's research aims, ensuring high biological relevance and excellent transfectability. With extensive experience in cell culture services, we support customers from routine culture conditions to specialized culture environments.
We operate advanced high-throughput phenotypic screening platforms, offering multiple endpoint assays including cell viability (MTT/MTS/XTT, ATP detection), apoptosis assays (Caspase activity, Annexin V/PI by flow cytometry), cell cycle analysis (PI staining by flow cytometry), and migration/invasion assays (Transwell, wound healing). For projects requiring more complex phenotype analysis, our high-content imaging system integrates automated microscopy with sophisticated image analysis technology to capture multi-parameter phenotypic data—such as cell number, morphology, cell cycle status, and apoptotic markers—with great resolution.
Experimental Design & Optimization
Before the project begins, our technical team collaborates closely with the client to design a customized experimental plan aligned with their research objectives. High-throughput screening is conducted in 96- or 384-well plates, with sufficient technical replicates and multiple control groups—negative, positive, and blank. Optimization of cell seeding density ensures cells remain in the logarithmic growth phase throughout the screening process.
Library Transfection & QC
Transfections use validated lipid-based reagents, such as Lipofectamine RNAiMAX, with optimized conditions tailored to each cell type. Library concentrations are maintained at nanomolar levels to maximize transfection efficiency while minimizing cytotoxicity. We also offer transfection efficiency assessment to ensure each experiment meets quality standards.
Phenotype Monitoring & Data Collection
Depending on the detection method chosen, we provide either real-time dynamic monitoring or endpoint assays. Real-time monitoring is implemented with RTCA systems, continuously recording cell index values post-transfection, generating complete growth curves. Endpoint assays are performed at designated time points post-transfection to obtain quantitative biological readouts.
Statistical Analysis
Data are rigorously analyzed using robust statistical methods. Raw data are normalized to the negative control mean. For RTCA data, key points such as relative cell index or area under the curve over specific time windows are calculated; for endpoint assays, ratios relative to negative controls are derived. Candidate hits are selected based on Z-score thresholds, depending on the experimental robustness and client tolerance for false positives.
Visualized Reporting
Clients receive a comprehensive data visualization package, including volcano plots displaying effect size versus significance, heatmaps illustrating hit miRNA impact on phenotypes, and time‐course trend charts for dynamic behavior. Each report includes a detailed description of methods, result interpretation, and recommendations for subsequent investigations.
Repeat Validation (Optional)
To ensure reliability and biological relevance, we validate hit miRNAs through repeat assays in the same cell line, dose–response testing, and cross-validation in additional cell models. For proliferation-related hits, further assays such as cell cycle, apoptosis, and colony formation are used to clarify their functional roles.
Creative Biogene's functional screening approach uncovers miRNAs with true biological roles, avoiding false positives common in expression-based selections. High-throughput design allows evaluation of hundreds of miRNAs in one project, greatly increasing efficiency. Real-time dynamic monitoring provides sensitive detection of nuanced phenotypic changes.
We enforce strict quality standards through carefully designed controls, multi-tier validation, and professional statistical analysis. Every project is backed by an experienced technical team and dedicated project management, ensuring smooth execution and reliable outcomes. We commit to delivering tailored service plans, timely communication, and expert support to help clients achieve breakthrough scientific insights.
If you're interested in functional miRNA screening or wish to learn more about our optional follow-up validation services, please contact us to discuss your project goals with our expert team.
Microarray-based miRNA Expression Profiling Service
MicroRNA Agomir/ Antagomir Synthesis Service
Q: Do you offer follow-up services to explore the mechanism of action of hit miRNAs identified from the screen?
A: Yes, we provide a full suite of optional mechanistic follow-up services to help elucidate the downstream targets and functional pathways of identified miRNAs:
Target Prediction & Pathway Analysis: We offer advanced bioinformatics support using tools like TargetScan, miRanda, PicTar, RNAhybrid, DIANA-microT, etc., to predict candidate target genes of hit miRNAs. Intersection across multiple algorithms enhances reliability. We also perform GO annotation and KEGG pathway enrichment analyses to focus on pathways most relevant to the phenotype.
Target Validation: We provide full validation services, including qRT-PCR for mRNA level changes, Western blot for protein expression, and dual-luciferase reporter assays using constructs containing wild-type and mutant 3'UTRs to directly confirm miRNA–target binding.
Functional Rescue Experiments: To confirm causality, we perform rescue assays: co-transfection of hit miRNA with ORF constructs of targets lacking miRNA binding sites to test phenotype reversal, or siRNA/shRNA knockdown of the target gene to mimic miRNA functional effects.
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