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

Di Yi Jun Yi Da Xue Xue Bao. 2004 Jan;24(1):35-8, 41.
[Article in Chinese]

Abstract

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.

Publication types

  • English Abstract
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basic Helix-Loop-Helix Transcription Factors
  • Bone Marrow Cells / cytology*
  • Cell Differentiation
  • DNA-Binding Proteins / antagonists & inhibitors*
  • DNA-Binding Proteins / genetics*
  • Dose-Response Relationship, Drug
  • Neurons / cytology*
  • RNA Interference*
  • RNA, Double-Stranded / pharmacology*
  • Rats
  • Rats, Sprague-Dawley
  • Repressor Proteins / antagonists & inhibitors*
  • Repressor Proteins / genetics*
  • Stem Cells / metabolism*
  • Transfection

Substances

  • Basic Helix-Loop-Helix Transcription Factors
  • DNA-Binding Proteins
  • Hes5 protein, rat
  • RNA, Double-Stranded
  • Repressor Proteins