Generation of MiRNA sponge constructs targeting multiple MiRNAs.

ABSTRACT

BACKGROUND: MicroRNAs(miRNAs) are key regulators of gene expression in plants, animals and some viruses. Hence, alteration of miRNA levels in cells or tissues is common for miRNA studies. Loss-of-function of miRNA can be achieved using antisense oligonucleotides, sponges and gene knockout models. METHODS: Here, we showed an efficient, rational and economical way to construct multi-targeted miRNA sponges with desired copies. Four copies of miRNA sponge are used as "building-blocks". RESULTS: These building-blocks, which can target same miRNA or different miRNAs, are linked together through ligation. Each time of ligation can double the number of sponge copies. CONCLUSIONS: In this way, we constructed lentivirus vectors harboring sponges targeting miR-21, miR-31 and miR-155 and the combination of two miRNA sponges can inhibit cancer cell growth significantly.