[Suppression of enhanced green fluorescent protein expression in cells by RNA interference].
OBJECTIVE
To evaluate the role of RNA interference (RNAi) in silencing the enhanced green fluorescent protein (eGFP) expression in 293T and Mel cells.


METHODS
Nested-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.


RESULTS
RNAi could effectively reduce more than 50 percent of eGFP expression in 293T cells as well as in Mel cells.


CONCLUSIONS
The RNAi vector constructed in this way paper can effectively inhibit eGFP expression in cells.
Xie
Shu-yang
SY
Institute of Medical Genetics, School of Medicine, Shanghai Jiaotong University, 200040 PR China.
Zhang
Jing-zhi
JZ
Huang
Shu-zhen
SZ
Ren
Zhao-rui
ZR
Zeng
Yi-tao
YT
D002460:Cell Line; D005434:Flow Cytometry; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006801:Humans; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins