The Nonviral Episomal Replicating Vector pEPI-1 Allows Long-Term Inhibition of bcr-abl Expression by shRNA

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The Nonviral Episomal Replicating Vector pEPI-1 Allows Long-Term Inhibition of bcr-abl Expression by shRNA
Apr 2005
Andreas C.W. Jenke, Tobias Eisenberger, Armin Baiker, Isa M. Stehle, Stefan Wirth, and Professor Hans Joachim Lipps
Human Gene Therapy
Human Gene Therapy
The Nonviral Episomal Replicating Vector pEPI-1 Allows Long-Term Inhibition of bcr-abl Expression by shRNA
Apr 2005, Vol. 16, No. 4: 533-539
Andreas C.W. Jenke
Children's Hospital, Helios Klinikum Wuppertal, 42283 Wuppertal, Germany.
Tobias Eisenberger
Institute of Cell Biology, University Witten/Herdecke, 58452 Witten, Germany.
Armin Baiker
Institute of Cell Biology, University Witten/Herdecke, 58452 Witten, Germany.
Isa M. Stehle
Institute of Cell Biology, University Witten/Herdecke, 58452 Witten, Germany.
Stefan Wirth
Children's Hospital, Helios Klinikum Wuppertal, 42283 Wuppertal, Germany.
Professor Hans Joachim Lipps
Institute of Cell Biology, University Witten/Herdecke, 58452 Witten, Germany.
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.

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