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Retroviral Infection of Murine Embryonic Stem Cell Derived Embryoid Body Cells for Analysis of Hematopoietic Differentiation
Published on: October 20, 2014
[Blocking with double- stranded RNA of the expression of Hes5 in rat bone marrow-derived neural stem cells]
Hai-yan Wang1, Ru-xiang Xu, Xiao-dan Jiang
1Institute of Neuromedicine of PLA, Zhujiang Hospital, First Military Medical University, Guangzhou 510282,China. haiyanwang838@msn.com
Objective:
To examine the efficiency of exogenous small double-stranded RNA (dsRNA) in knocking down the gene expression at the post-transcription level, and investigate the factors that may influence the transfection.
Method:
The bone marrow stromal cells of SD rat were separated and cultured in vitro, followed by induction of the cells to evolve into neural stem cells using special culture medium prepared by our laboratory. Synthetic dsRNA was then transferred into the cells at varied concentrations, and the results were analyzed by Western blotting.
Results:
The concentrations ranging from 200 to 300 nmol/L were optimal for specifically blocking the expression of Hes5, whereas the suitable concentrations for the cell survival were between 50 and 200 nmol/L.
Conclusion:
dsRNA is capable of triggering RNA interference in neural stem cells, and at appropriate concentration, it may specifically and effectively knock down endogenous gene expression without sacrificing the viability of the cells.
