Rapad Mirna Adenoviral Assay Kit

Background: MicroRNAs (miRNAs) are 18-24 nucleotide RNA molecules that regulate the stability or translational efficiency of target mRNAs. These regulatory RNAs function by acting as sequence-specific guides which recruit a large protein complex known as the RNA-induced silencing complex (RISC) to target mRNAs which are subsequently silenced. Diverse functions have been attributed to miRNAs including the regulation of cellular differentiation, proliferation, and apoptosis. Moreover, significant evidence has accumulated implicating a fundamental role for miRNAs in the development of cancer. miRNAs are initially transcribed as long precursor transcripts known as primary microRNAs (pri-miRNAs). Within these transcripts, the mature miRNA sequences are found in ~60-80 nucleotide hairpin structures. Mature miRNAs are generated from pri-miRNAs by sequential processing (Figure 1). Pri-miRNAs are initially recognized in the nucleus by the microprocessor complex which includes as core components the RNase-III enzyme Drosha and its obligate partner DGCR8. This complex excises the hairpin structure containing the mature miRNA sequence. The liberated hairpins, referred to as precursor miRNAs (pre-miRNAs), are recognized by the nuclear export factor exportin 5 which transports them to the cytoplasm. There, the RNase-III enzyme Dicer performs a second cleavage to generate a double-stranded 18-24 nucleotide RNA molecule. The RISC then associates with this RNA duplex and unwinds it. Generally, only one strand is stably incorporated into the RISC; the other is discarded and rapidly degraded. miRNAs guide the RISC to target messages that are subsequently cleaved or translationally silenced (Figure 1). Recombinant adenoviruses have tremendous potential in both research and therapeutic applications. There are numerous advantages they provide when introducing genetic material into host cells. The permissive host cell range is very wide. The virus has been used to infect many mammalian cell types (both replicative and non-replicative) for high expression of the recombinant protein. Recombinant adenoviruses are especially useful for gene transfer and protein expression in cell lines that have low transfection efficiency with liposome. After entering cells, the virus remains epichromosomal (i.e. does not integrate into the host chromosome so does not activate or inactivate host genes). Recently, recombinant adenoviruses have been used to deliver RNAi into cells. RAPAd Adenoviral Expression System provides a much faster and safer method to generate RCA-free recombinant adenovirus at high titer. The RAPAd system uses a novel Ad backbone devoid of the left-hand ITR, the packaging signal and E1 sequences. There is no need to perform the bacterial in vitro homologous recombination (pAdEasy method), and also the multiple plaque isolations (standard homologous recombination method in packaging cell line). The RAPAd system allows for generation of a recombinant virus within 2 weeks and the virus produced contained virtually no contaminating E1a sequences or replication-competent virus (RCA). RAPAd miRNA Adenoviral Expression System is designed to rapidly produce recombinant adenovirus that expresses an individual miRNA precursor in its native context while preserving putative hairpin structures to ensure biologically relevant interactions with endogenous processing machinery and regulatory partners, leading to properly cleaved microRNAs. Individual miRNA precursor from any species can be cloned between BamHI and Nhe I sites (Figure 2).

Small volumes of RAPAd miRNA Adenoviral assay kit vial(s) may occasionally become entrapped in the seal of the product vial during shipment and storage. If necessary, briefly centrifuge the vial on a tabletop centrifuge to dislodge any liquid in the container`s cap. Certain products may require to ship with dry ice and additional dry ice fee may apply.

MBS169399 is an assay kit targeting RAPAd miRNA Adenoviral. Also known as RAPAd miRNA Adenoviral Expression System.