[Blocking with double- stranded RNA of the expression of Hes5 in rat bone marrow-derived neural stem cells].
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.


METHODS
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.


CONCLUSIONS
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.
Wang
Hai-yan
HY
Institute of Neuromedicine of PLA, Zhujiang Hospital, First Military Medical University, Guangzhou 510282,China. haiyanwang838@msn.com
Xu
Ru-xiang
RX
Jiang
Xiao-dan
XD
Luo
Shen-qiu
SQ
D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D009474:Neurons / Q000166:cytology*; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012097:Repressor Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics*; D013234:Stem Cells / Q000378:metabolism*; D014162:Transfection
D051792:Basic Helix-Loop-Helix Proteins; D004268:DNA-Binding Proteins; D012330:RNA, Double-Stranded; D012097:Repressor Proteins; C489686:Hes5 protein, rat