Small RNA is a large class of regulatory molecules, including miRNA, siRNA, snoRNA, piRNA, and rasiRNA. Among them, miRNAs are endogenous small RNA molecules of about 20-24 nucleotides in length, which are involved in regulating the growth and development of organisms and the occurrence of diseases in cells mainly through degrading mRNAs, inhibiting translation, and regulating the formation of heterochromatin. Each miRNA can have multiple target genes, and several miRNAs can target the same gene, thus forming a complex regulatory network. Lifeasible can provide various research solutions by combining bioinformatics research tools to help you decipher the critical role of miRNAs in regulating gene expression, cell cycle, and developmental timeline of organisms.

Bioinformatics prediction of target genes

We used a bioinformatics approach to predict the target gene loci of miRNAs, i.e., the site of action silenced by the miRNA gene.

Confirmation of target gene prediction results

After performing target site prediction, we can identify the target gene locus by miRNA pulldown method. We provide the following three commonly used pulldown methods for you to choose from.

• Biotinylated miRNA pull-down. Our service steps include biotinylated miRNA transfection into cells, cells lysed after incubation, and streptavidin-coated magnetic beads adsorbed to screen for miRNA and its effector mRNA.
• Labeled microRNA pull-down assay (LAMP). We obtained co-precipitated mRNAs by incubating pre-miRNA oligonucleotides labeled with DIG mixed with cell extracts and doing immunoprecipitation (IP) with anti-DIG antibodies.
• Ribonucleoprotein immunoprecipitation followed by microarray chip analysis (RIP-Chip, RIP-seq). We performed a microarray analysis of the mRNAs obtained by transfecting cells with synthetic miRNAs, lysing the cells after incubation, and immunoprecipitating RISC with specific anti-AGO2 antibodies.

Direct functional confirmation of miRNAs

We help you verify whether miRNAs can specifically bind to target mRNAs and inhibit their expression through experimental methods.

• 3'-UTR reporter assay

We insert the target gene to be confirmed into the downstream 3'-UTR region of a reporter gene, such as luciferase CDS, clone it into a vector, transfect the cells with the vector, and then treat the cells with miRNA. If the target gene contains a target site, the transcription of luciferase is inhibited from fluorescing.

• Up- and down-regulation of miRNAs

We enhance or inhibit the inhibitory effect of miRNAs by artificially synthesizing miRNA mimics or miRNA inhibitors and comparing them with the control.

• Site-directed mutagenesis

In addition, Lifeasible can provide you with solutions for overall functional validation and quantitative and localization analysis of miRNAs. We explore the up-/down-regulation of miRNAs through Western Blot or other biological pathway analysis experiments to resolve how miRNAs ultimately affect cells, diseases, and other conditions. Through RT-qPCR, in situ hybridization, microarray, and other experiments, we can determine the exact expression of miRNAs and their locations to complement their mechanistic studies. Please feel free to contact us for the most comprehensive miRNA analysis solutions.

*If your organization requires the signing of a confidentiality agreement, please contact us by email.

For research use only, not intended for any clinical use.

Related Services

• Small RNA Library Analysis
• Non-Coding RNA Gene Identification
• Theoretical Predictions of Non-Coding RNA Genes
• MicroRNA Detection
• Small Interfering RNA
• RNA Modification Detection