A miR-21 hairpin structure-based gene knockdown vector

Junming Yue, Yi Sheng, Aixia Ren, Sravya Penmatsa

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

RNA interference (RNAi) 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. The miR-21 hairpin-based gene knockdown vector will provide a new genetic tool for gene functional studies in vitro and in vivo.

Original languageEnglish (US)
Pages (from-to)667-672
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume394
Issue number3
DOIs
StatePublished - Apr 9 2010

Fingerprint

Gene Knockdown Techniques
Small Interfering RNA
Genes
Lamin Type A
MicroRNAs
Reverse Genetics
RNA Interference
Reporter Genes
Fluorescence
Nucleotides
RNA

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

A miR-21 hairpin structure-based gene knockdown vector. / Yue, Junming; Sheng, Yi; Ren, Aixia; Penmatsa, Sravya.

In: Biochemical and Biophysical Research Communications, Vol. 394, No. 3, 09.04.2010, p. 667-672.

Research output: Contribution to journalArticle

Yue, Junming ; Sheng, Yi ; Ren, Aixia ; Penmatsa, Sravya. / A miR-21 hairpin structure-based gene knockdown vector. In: Biochemical and Biophysical Research Communications. 2010 ; Vol. 394, No. 3. pp. 667-672.
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