Construction and functional characterization of alentiviralvector mediating RNA interference of DEK gene
10.7644/j.issn.1674-9960.2015.07.003
- VernacularTitle:DEK基因RNAi慢病毒表达载体的构建及其功能鉴定
- Author:
Jie ZHU
;
Jie LIU
;
Lihua DING
;
Ronghua HE
;
Yanan ZHANG
;
Zhida CHEN
;
Xiaoli LUO
;
Qinong YE
- Publication Type:Journal Article
- Keywords:
DEK;
RNA interference( RNAi);
lentiviral vector;
breast neoplasms;
ZR75-1;
gene expression;
plasmids;
transfection
- From:Military Medical Sciences
2015;(7):499-503
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct the lentiviral vector of RNA interference(RNAi) for DEK,and to detect its effect on breast cancer cell growth.Methods The DEK siRNA was designed and constructed based on DEK sequence using a lentiviral vector.The lentivirul vector containing DEK siRNA was named PSIH-H1-DEK as confirmed by PCR and sequenceing.PSIH-H1-DEK was then packaged with accessory plasmids into lentivirus in 293T cells and selected for 2 weeks with puromycin ( puro ) before the mixed colonies stably expressing DEK siRNA were obtained and the DEK expression was detected by real time PCR( RT-PCR) and Western blotting.The effect of DEK siRNA on ZR75-1 cell growth was determined by cell counting kit.Results Western blot and RT-PCR showed that PSIH-H1-DEK siRNA could suppress DEK gene expression.Suppression of DEK could markedly inhibit the growth of ZR75-1 cells.Conclusion The lentivirus-mediated DEK siRNA is obtained,which will facilitate further research on DEK function in breast cancer development.
