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. 2013 Apr 16;21(6):1259–1269. doi: 10.1038/mt.2013.65

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Copyright © 2013 The American Society of Gene & Cell Therapy

This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 3.0 License. To view a copy of this license, visit http://creativecommons.org/licenses/by-nc-nd/3.0/

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Protein kinase C (PKC) activators (PMA and bryostatin) significantly enhanced CCR5-ZFN activity in CD34⁺ hematopoietic stem/progenitor cells (HSPC). Adult HSPCs were stimulated with SFT in SCGM medium overnight followed by PMA or bryostatin treatment for 30 minutes before CCR5-ZFN transduction (multiplicity of infection (MOI) 150). The percent CCR5 disruption was obtained by surveyor nuclease assay from transduced cells (MOI 150) without PKC activator treatment (left), with 20 ng/ml PMA treatment (middle), and 10 nmol/l bryostatin treatment (right). Percent of CCR5 disruption is indicated under each lane. PMA, phorbol 12-myristate 13-acetate; ZFN, zinc finger nuclease.