ural stem cells, which grow as neurospheres in the culture medium, a 50% reduction in protein synthesis of the target gene was observed. The lower efficiency of gene silencing in these primary culture cells can be attributed to inefficient transfection observed with cells grown in suspension and poor transport of the siRNAs to the core of the sphere. Cells transfected with Oligofectamine alone or single-stranded RNA as controls did not show altered levels of the targeted protein. Moreover, nonspecific RNAi effects, such as activation of the PKR pathway, were not observed. In some cases, when growth factors were removed from the culture medium, differentiation of the primary neural stem cells, which normally give rise to neuronal and glial cells, was altered in the siRNA treated cells. siRNA-mediated gene silencing is thus proving to be a powerful tool for the study of differentiation of mouse neuronal primary cells.

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References

1. Zamore PD, Tuschl T, Sharp Pa, Bartel DP. (2000) RNAi:
double stranded RNA directs the ATP-dependent cleavage of mRNA at 21 to 23 nucleotide intervals. Cell. 101(1): 2533.

2. Elbashir SM, Lendeckel W, and Tusch T (2001) RNA interference is mediated by 21- and 22- nucleotide RNAs. Genes Dev. 15(2): 188200.

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