A miR-21 hairpin structure-based gene knockdown vector

作者：Yue Junming^*; Sheng Yi; Ren Aixia; Penmatsa Sravya

来源：Biochemical and Biophysical Research Communications, 2010, 394(3): 667-672.

DOI：10.1016/j.bbrc.2010.03.047

• miRNA
• miR-21
• Gene knockdown
• Lentiviral vector

RNA interference (RNA1) is widely used to study gene functions as a reverse genetic means from first-generation siRNA to second-generation short hairpin RNA (shRNA) or the newly developed microRNA (shRNA-miR) Here we report a gene knockdown vector system based on the mouse miR-21 hairpin structure In this system, the pre-miRNA hairpin of the miR-21 gene was modified by replacing the 22-nucleotide mature sequence with shRNA sequences that target genes of interest, flanked by 160-bp upstream and 65-bp downstream sequences of the mouse pre-miR-21 We tested this system by knocking down the enhanced green fluorescence protein (EGFP) reporter gene using different vectors, in which shRNA-miR was driven by the polymerase II (pol II) promoter We found that miR-21 hairpin-based shRNA-miR can be directly placed under pol II promoter, like UbC or CMV promoter to knockdown the gene of interest To facilitate the wide application of the miR-21 hairpin-based gene knockdown system, we further knocked down the endogenous gene lamin (A/C), which showed that endogenous lamin A/C expression can be efficiently silenced using the miR-21 hairpin-based lentiviral vector.

• 出版日期2010-4-9