MiRNA Target Reporter Vectors

MicroRNAs regulate gene expression by targeting messenger RNAs at specific sites to induce the cleavage of the targets or inhibit translation. The interaction of a miRNA with its targets is known mediated by RNA-induced silencing complex (RISC). RISC-mediated interaction between a mature miRNA and its binding site depends on sequence complementarities between miRNA and its targets and the total number of binding sites in a given 3’UTR. Numerous important regulatory genes have been found to be regulated by miRNAs, such as Ras by let-7, Bcl-2 by miR-15 and miR-16, ERa by miR206, TPM1 by miR-21, and PTEN by miR-19a. Luciferase reporter-based cellular assays are often employed to monitor the regulation of these genes by miRNAs.

A. Benefits:

MiRNA Target Luciferase Reporter Vectors offer the following benefits:

• Functional analysis - It provides functional assays to monitor miRNA-mediated regulation of taregt genes in cells.

• Quantitative assay - MiRNA-mediated regulation can be quantitatively measured through repression of firefly luciferase gene expression.

• Stable cell lines - The vectors can be used for establishing stable cells expressing the reporters.

B. Principle of the technology

pLuc-3UTR reporter vectors are a series of firefly luciferase-based reporter constructs for monitoring miRNA-mediated regulation of target genes in cells. Each vector contains the CMV promoter, firefly luciferase gene, the 1kb 3’UTR of a target gene, and a SV40 terminator sequence. When a miRNA is expressed and binds to 3'UTR, it results in repression of luciferase gene expression. Therefore, miRNA-mediated regulation of a specific target can be monitored through luciferase activity. When two samples are compared by measuring the activities of luciferase, the difference can be identified and the regulation dissected.