[Construction and selection of RNAi plasmid vector for suppressing GATA-3 gene expression].
GATA-3 plays a central role in the Th2-mediated immunoreaction. This study was aimed to construct and select plasmid vectors of siRNA which can effectively and specifically suppress the gene expression of GATA-3. Plasmid including PSi338, PSi717 and PSi1232 were designed and constructed for GATA-3 regarded as target gene and transfected into murine P815 cells. Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot were used to detect the inhibition of GATA-3 mRNA as well as its protein in P815 cells. The results demonstrated that the expressions of mRNA and protein of GATA-3 in P815 cells were inhibited significantly by both of PSi338 and PSi717. It is concluded that PSi338 and PSi717 siRNA plasmid vectors have been successfully constructed, and both vectors are effective and specific siRNA plasmids for suppressing GATA-3 gene expression.
Zhou
Bing
B
Department of Otorhinolaryngology, Head and Neck Surgery, Tongren Hospital, Capital University of Medical Science, Beijing 1000730, China. entzhou@263.net
Zheng
Ming
M
Wang
Yan
Y
DU
Zhi-Yan
ZY
Wang
Tong
T
D000818:Animals; D015536:Down-Regulation; D050990:GATA3 Transcription Factor / Q000378:metabolism*; D005822:Genetic Vectors; D051379:Mice; D010957:Plasmids; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics*; D018418:Th2 Cells / Q000276:immunology*; D014162:Transfection
D050990:GATA3 Transcription Factor; C494748:Gata3 protein, mouse; D012333:RNA, Messenger; D034741:RNA, Small Interfering