Interference of OX40 gene expression induced by small interfering RNA
- VernacularTitle:小干扰RNA对大鼠OX40基因表达的沉默作用
- Author:
Renpin XIA
;
Shichun LU
;
Jushan WU
;
Ning LI
;
Jing ZHENG
- Publication Type:Journal Article
- Keywords:
OX40;
Gene silence;
Small interfering RNA
- From:
Chinese Journal of Immunology
1986;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the specific interference of OX40 gene expression induced by RNAi technique in 293T cell lines transfected with rat OX40 gene.Methods:293T cells were transfected with recombined plasmid pEGFP-N1-GFP/OX40,and the positive cell clones were selected by fluorescence protein observation and RT-PCR.One specific dicer siRNA targeted to OX40 mRNA was designed and synthesized,which shared no homology with exons of known human gene.Quantitative real-time PCR was performed to measure the inhibitory rate of target gene expression by comparing OX40 mRNA concentrations before and after siRNA transfection.Results:10 nmol/L siRNA-OX40 elicited the highest level of gene silence in 293T cells which was transfected with siRNA after 48 h (68.3?8.7)%);The time of maximal inhibitory effect was at 48-72 h [(61.7?8.4)%,(39.6?5.6)%].Conclusion:The exogenous OX40 expression can be significantly inhibited by treatment with specific siRNA in a dose and time -dependent manner in 293T cells,which may provide a useful profile for further investigation of inhibition of OX40 protein,and a promising control approach for preventing immune reaction.
