Suppression of enhanced green fluorescent protein expression in cells by RNA interference.
- Author:
Shu-yang XIE
1
;
Jing-zhi ZHANG
;
Shu-zhen HUANG
;
Zhao-rui REN
;
Yi-tao ZENG
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line; Flow Cytometry; Genetic Vectors; genetics; Green Fluorescent Proteins; genetics; Humans; RNA Interference; Reverse Transcriptase Polymerase Chain Reaction
- From: Chinese Journal of Medical Genetics 2005;22(4):431-434
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo evaluate the role of RNA interference (RNAi) in silencing the enhanced green fluorescent protein (eGFP) expression in 293T and Mel cells.
METHODSNested-PCR was used to amplify H1 promoter from human 293T cells for driving RNAi synthesis. RNAi vectors (TR1) for silencing the eGFP expression was constructed. The eGFP vector and RNAi vector (TR1) were then co-transfected into the 293T and Mel cells, in which the silencing effect on eGFP expression was investigated by fluorescence microscopy, reverse transcription-PCR(RT-PCR), fluorescence-assited cell sorting(FACS) analysis and real-time RT-PCR.
RESULTSRNAi could effectively reduce more than 50 percent of eGFP expression in 293T cells as well as in Mel cells.
CONCLUSIONThe RNAi vector constructed in this way paper can effectively inhibit eGFP expression in cells.
