miRNA is an important factor for regulating gene expression, and the regulation is reflected in many aspects, for example, one miRNA may regulate multiple target genes, and one target gene can be regulated by a number of miRNA , so it may produce a complex and large regulatory networks. RiboBio can provide you with miRNA mimics library and miRNA inhibitor library, so that you can study the miRNA which play an important role in this regulatory network from a variety of point of view.
The functional analysis of miRNA is similar to genes. Over-expression and specific inhibition are two common effective research methods.
Over-expression: Transfect the miRNA mimics into cells and then you can achieve transient over-expression of the miRNA. miRNA agomir can realize long time miRNA over-expression effects. Normal cell experiments use miRNA mimics.
Specific inhibition: Transfect the miRNA inhibitor into cells to test the specific inhibitory function of miRNA. The chemical modification of miRNA antagomir enhances its stability, this is widely used in animal experiments.
miRNA research is usually carried out in cell or in vivo, and it is always combined with cell phenotype (such as cell proliferation, apoptosis, cell migration and so on) and gene expression analysis.
RiboBio has first-class miRNA specialty products and strong technical advantages. We can provide you with a reliable miRNA functional analysis service.
You just need to inform the miRNA name and cell lines.
1) miRNA proliferation screening service
Cell proliferation is an important life characteristics of organisms, and cancer cells can proliferate nearly unlimited especially.EdU detection method is the latest detection for cell proliferation. The structure of EdU is similar to T bases, so EdU can replace the T base to insert in the copying cells. To detect DNA replication activity based on the specific reaction of EdU and Apollo ® fluorescent dye, we can obtain accurate cell proliferation by detecting the EdU mark.
Just as the example of miR-x, miR-y research, 24h after the micrON ™ of miRNA mimics library transfecting into A549 cell line, special function miRNA can be screening by EdU cell proliferation detection method. miR-x obviously inhibits cell proliferation while miR-y obviously promotes cell proliferation.
Figure 1. Using miRNA library for high content screening experiments and the detection results show that miR-x and miR-y have significant effect on cell proliferation.
A: the overall picture, 4X4; B: partial decomposed picture; C: miR-x obviously inhibits cell proliferation while miR-y obviously promotes cell proliferation.
2)miRNA signal pathway screening service
Figure 2. Using miRNA inhibitor library to screen miRNA which has special effects on NF-kB reporter gene.
Both micrOFF™ miRNA inhibitor library and NF-kB reporter gene plasmid vector were co-transfected into A549 cells, and to study the impact that miRNA inhibitor has on NF-kB signaling pathway by detecting changes of luciferase activity 48 h later.