Construction of TDRG1 shRNA expression vector and interfering effect of TDRG1 shRNA expression vector on NTERA-2 cells.
10.3969/j.issn.1672-7347.2012.10.002
- Author:
Shenglin PENG
1
;
Jianfu YANG
;
Houyang CHEN
;
Xiaoliang GUO
;
Dongjie LI
;
Huabo ZHOU
;
Yu GAN
;
Xianzhen JIANG
;
Yuxin TANG
Author Information
1. Department of Urology, Third Xiangya Hospital, Central South University, Changsha, China.
- Publication Type:Journal Article
- MeSH:
Cell Line, Tumor;
Genetic Vectors;
Humans;
RNA Interference;
RNA, Messenger;
RNA, Small Interfering;
Transfection
- From:
Journal of Central South University(Medical Sciences)
2012;37(10):979-982
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To construct short hairpin RNA interfering expression vector of TDRG1,and detect the specific interfering effect of TDRG1-shRNA expression vector on NTERA-2 cells.
METHODS:Oligos for short hairpin RNA targefing for TDRG1 were designed and connected to the expression vector pGPU6/GFP/Neo to construct the TDRG1 shRNA expression vector. The recombinant plasmid TDRG1-shRNA486, TDRG1-shRNA738, TDRG1-shRNA921 and lipofectamine ™2000 were used to generate and transfect shRNA into NTERA-2 cells. Expression of TDRG1 mRNA was assayed by RT-PCR.
RESULTS:TDRG1-shRNA expression vector was successfully constructed. TDRG1-shRNA486 was more effective in the suppression of TDRG1 with significant reduction of TDRG1 mRNA.
CONCLUSION:TDRG1-shRNA can interfere the expression of TDRG1 in NTERA-2 cells.