Screening of siRNA expression plasmid by nondenaturing polyacrylamide gel electrophoresis

Objective To screen TGF-β_1 specific siRNA expression plasmid by nondenaturing polyacrylamide gel electrophoresis.Methods Human lung adenocarcinoma A549 as tumor cell model,siRNA sequences were selected by Angela principle and siRNA plasmids were screened by nondenaturing polyacrylamide gel electrophoresis.Then A549 cells were transfected with the reformed plasmids.The expression level of TGF-β_1 were detected by indirect immunofluorescence assay(IFA) and Western blotting.Results POligo 1210,pOligo 1216,pOligo 1363 TGF-β_1were constructed successfully.The insert sequence can be distinguished in 12% nondenaturing polyacrylamide gel electrophoresis.SiRNA plasmids were screened by nondenaturing polyacrylamide gel electrophoresis successfully.TGF-β_1 expression can be inhibited with three plasmids ranged from 50% to 83.2%.Conclusion TGF-β_1 specific siRNA plasmids were screened by nondenaturing polyacrylamide gel electrophoresis successfully.It will be helpful to the study of TGF-β_1 in tumor cells.