MicroRNAs (miRNAs) are naturally occurring 21-23 nt noncoding RNAs that are involved in many biological processes and disease development. They have been found in all species and regulates the translation and degradation of its target messenger RNA by binding to complementary regions in mRNA.
Creative Biogene provides a comprehensive miRNA expression plasmids covering human, mouse, and rat genomes to detect the effects of miRNAs on cell functions and phenotypes. These vectors provide powerful miRNA expression and allow scientists to validate and track the expression using fluorescent protein markers. In addition, we also offer cloning services to design the most suitable custom products according to specific needs.
The miRNA clone insert of Creative Biogene consists of pri-mir (60-70nts) and 250-300nts genomic sequences flanking it. In general, flanking sequences are required for correct pri-mir expression and mature microRNA processing. To test miRNA expression, isolate small RNAs from transfected cells. RT-PCR and qPCR are then performed using specific miRNA primers to measure miRNA expression. Creative Biogene randomly selected three overexpression plasmids and used qPCR to demonstrate the expected overexpression of miRNA in all cases.
And all miRNA clones were sequenced to ensure that the pre-mir sequence matched the reference sequence in miRBase. In terms of biological polymorphisms, the exact flanking sequence of a miRNA clone may differ from the NCBI reference, which will not affect the function of the mature miRNA. In Creative Biogene, we monitor the transfection of tGFP signals. The PCMV-mir vector contains an IREStGFP reporter gene, which is driven by the SV40 promoter and located downstream of the neomycin gene. Independent tGFP and pre-mir expression from different promoters are designed to minimize interference between the two expression cassettes, so tGFP can truly reflect transfection efficiency.
Each vector is equipped with a powerful CMV promoter, a fluorescent tag gene, and a selection marker. Clone the miRNA sequence of interest to the downstream of the fluorescent protein gene. And then, the expression of miRNA can be analyzed by observing the expression level of the fluorescent protein. Using these vectors, researchers can clone and express miRNAs needed, then select, classify and/or visualize the cells that express them.
The Features make Creative Biogene distinctive:
.Offer >30, 000 miRNA expression plasmids
.Genome-wide miRNA coverage of multi-species (including humans, mice, and rats)
.Provide a variety of fluorescent markers for use as a reporter for transfection monitoring – GFP, mCherry, Luc2, ZsGreen1, etc.
.Provide a variety of selectable markers to create stable cell lines – Neomycin, Puromycin, Hygromycin, etc.
Creative Biogene miRNA expression plasmids can help you
.Efficient transfection in a variety of cell types including dividing, non-dividing cells, adult and embryonic stem cells, and iPS cells, etc.
.Express your selected miRNAs in vivo and in vitro to reduce target gene expression
.Generate stable cell lines for long-term expression of miRNAs
