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ABSTRACT RNA interference (RNAi) has recently been used for sequence-specific gene silencing of disease-related genes including oncogenes in hematopoietic cells. To characterize its

potential therapeutic value, we analyzed different modes to activate RNAi as well as some pharmacokinetic aspects of gene silencing in bcr-abl+ cells. Using lentiviral gene transfer of

transcription cassettes for anti-bcr-abl shRNAs and red fluorescence protein (RFP) as a quantitative reporter, we demonstrate that stable but not transient RNAi can efficiently deplete

bcr-abl+ K562 and murine TonB cells from suspension cultures. Importantly, depletion of bcr-abl+ cells depends on the dose of lentivirus used for transduction and correlates with the

RFP-expression level of transduced target cells: RFP-high K562 cells are eradicated, whereas RFP-low or -intermediate cells may recover after prolonged cell culture. Interestingly, these

cells still show reduced bcr-abl mRNA levels, aberrant proliferation kinetics, and enhanced sensitivity to the Bcr-Abl-kinase inhibitor STI571. Quantitative PCR from genomic DNA suggests

that more than three lentiviral integrations are required for effective depletion of K562 cells. Finally, we demonstrate that lentivirus-mediated anti-bcr-abl RNAi can inhibit colony

formation of primary CD34+ cells from chronic myeloid leukemia patients. These data demonstrate dose-dependent gene silencing by lentivirus-mediated RNAi in bcr-abl+ cells and suggest that

stable RNAi may indeed be therapeutically useful in primary hematopoietic cells. Access through your institution Buy or subscribe This is a preview of subscription content, access via your

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* Learn about institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS ERYTHROID LINEAGE-SPECIFIC LENTIVIRAL RNAI VECTORS SUITABLE FOR

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This work was supported in part by grants of the HW and J Hector-Stiftung, the Wilhelm Sander-Stiftung, and the ‘Deutsche Forschungsgemeinschaft’ (SFB 566). We thank George Daley (MIT,

Cambridge) for providing us with the TonB cell line used in this study. AUTHOR INFORMATION AUTHORS AND AFFILIATIONS * Department Hematology and Oncology, Hannover Medical School, Hannover,

Germany M Scherr, K Battmer, A Ganser & M Eder * III Med. Clinic, University Hospital of Mannheim, University of Heidelberg, Mannheim, Germany B Schultheis Authors * M Scherr View author

publications You can also search for this author inPubMed Google Scholar * K Battmer View author publications You can also search for this author inPubMed Google Scholar * B Schultheis View

author publications You can also search for this author inPubMed Google Scholar * A Ganser View author publications You can also search for this author inPubMed Google Scholar * M Eder View

author publications You can also search for this author inPubMed Google Scholar RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS ARTICLE CITE THIS ARTICLE Scherr, M., Battmer, K.,

Schultheis, B. _et al._ Stable RNA interference (RNAi) as an option for anti-bcr-abl therapy. _Gene Ther_ 12, 12–21 (2005). https://doi.org/10.1038/sj.gt.3302328 Download citation *

Received: 01 April 2004 * Accepted: 30 May 2004 * Published: 16 December 2004 * Issue Date: 01 January 2005 * DOI: https://doi.org/10.1038/sj.gt.3302328 SHARE THIS ARTICLE Anyone you share

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Nature SharedIt content-sharing initiative KEYWORDS * RNA interference * siRNA * lentivirus * gene transfer * bcr-abl