THE SILENCE OF SHORT HAIRPIN RNAs INDUCED Smad2 IN NIH/3T3 FIBROBLAST CELLS
- VernacularTitle:shRNAs介导的Smad2基因在小鼠成纤维细胞NIH/3T3中表达的沉默
- Author:
Rong ZHENG
;
Qi XIONG
;
Siwen JIANG
;
Bo ZUO
;
Fenge LI
;
Dequan XU
;
Zhuqing REN
;
Yuanzhu XIONG
- Publication Type:Journal Article
- Keywords:
RNAi;
Gene silence;
shRNA;
TGF-?;
Smad2;
Western blotting;
Mouse
- From:
Acta Anatomica Sinica
2002;0(05):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct five shRNA-expression plasmids and to investigate the expression of Smad2 in TGF-?/ Smads signal transduction treated with shRNA-expression plasmid.Methods Five shRNA-Smad2 DNA sequences from mRNA sequence of mouse Smad2 gene were designed and synthesized.DNA oligonucleotides encoding an appropriate shRNA were inserted to shRNA expression vector respectively.Five shRNA-Smad2 expression plasmids were obtained and then transfected into NIH/3T3 cells.The suppressed expression of Smad2 was assessed by RT-PCR and Western-blotting.Results The shRNA-expression plasmid numbered 2.4 could markedly reduce the expression of Smad2.The suppression effect of the RNAi-pool composed of four different plasmids was more obvious than that of any single.Conclusion The shRNA-expression plasmids were successfully constructed,which could specifically and effectively suppress the expression of Smad2.The method of using a mixture of RNAi plasmids to improve the RNAi efficiency was established.
