Construction and screening of RNA interference vector against A20 gene

Objective Our previous study has shown that A20 was over-expressed in both human glioma tissues and cell lines. The objective of this study is to construct the RNA interference vector of the A20 and to observe its inhibitory effect on A20 expression in U251 cell.Methods Three short hairpin RNA (shRNA) fragments targeting A20 were designed and introduced into pSilencer3.1 plasmid to construct eukaryotic siRNA vectors pSilencer3.1-H1-A20R. U251 cells were transfected with three different siRNA vectors by liposome method. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression of A20 in the transfected cells.Results Three different siRNA vectors were constructed successfully. One of vectors can significantly reduce the expression of A20 both at mRNA and protein level.Conclusion The siRNA vector targeting A20 is obtained, which provides the premise for further studying the role of A20 in proliferation, angiogenesis and apoptosis of the glioma cells.