Suppression of enhanced green fluorescent protein expression in cells by RNA interference / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
;
(6): 431-434, 2005.
Article
in Chinese
| WPRIM
| ID: wpr-280034
ABSTRACT
<p><b>OBJECTIVE</b>To evaluate the role of RNA interference (RNAi) in silencing the enhanced green fluorescent protein (eGFP) expression in 293T and Mel cells.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>RNAi could effectively reduce more than 50 percent of eGFP expression in 293T cells as well as in Mel cells.</p><p><b>CONCLUSION</b>The RNAi vector constructed in this way paper can effectively inhibit eGFP expression in cells.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Cell Line
/
Reverse Transcriptase Polymerase Chain Reaction
/
RNA Interference
/
Green Fluorescent Proteins
/
Flow Cytometry
/
Genetic Vectors
/
Genetics
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Medical Genetics
Year:
2005
Type:
Article
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