alexa The nonviral episomal replicating vector pEPI-1 allows long-term inhibition of bcr-abl expression by shRNA.
Molecular Biology

Molecular Biology

Journal of Cell Science & Therapy

Author(s): Jenke AC, Eisenberger T, Baiker A, Stehle IM, Wirth S,

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Abstract The inhibition of gene expression by RNA interference harbors a high potential for application in the therapy of human diseases. However, while exogenous application of siRNAs efficiently inhibits gene expression, these effects are only transient in mammalian cells. We designed a short hairpin RNA-expression cassette to target the bcr-abl oncogene that was then introduced into the nonviral vector system pEPI-1, which replicates episomally in the absence of selection in the bcr-abl-positive cell line K562. Forty-two days after transfection the bcr-abl- but not the cytokine-dependent growth rate was found to be drastically reduced in K562 cells. Western analysis revealed a more than 90\% reduction in the expression of the fusion protein bcr-abl while the expression of the bcr protein remained unaffected. In addition, we show that the level of bcr-abl mRNA was specifically reduced in these cells for more than 90\%. These results demonstrate that the vector system pEPI-1 allows specific and efficient long term gene suppression by using a short hairpin RNA transcription unit. This article was published in Hum Gene Ther and referenced in Journal of Cell Science & Therapy

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