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. 2018 Aug 28;9:1949. doi: 10.3389/fimmu.2018.01949

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Copyright © 2018 Hammerschmidt, Werth, Rothe, Galla, Permanyer, Patzer, Bubke, Frenk, Selich, Lange, Schambach, Bošnjak and Förster.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The unlimited proliferative capacity of Cas9-Hoxb8 cells allows the consecutive knockout of multiple genes.Ccr7^−/− Cas9-Hoxb8 cells were transduced with a Cerulean- and CXCR4-gRNA-encoding lentivirus. Transduced cells as well as control Cas9-Hoxb8 cells were subsequently differentiated to mature DCs in the presence of GM-CSF followed by the treatment with LPS. DCs were analyzed by flow cytometry for their expression of CD80, CCR7, and CXCR4. Gray curves depict isotype controls. Data are representative of two independent experiments.