Abstract
Hoxb8 cells are immortalized myeloid progenitors that maintain their multipotent potential and can be differentiated into neutrophils. Genetic modification of Hoxb8 cells can be used as a model system for the functional analysis of regulators of neutrophil maturation and effector functions, such as transcription factors. Here we describe the generation of transcription factor (TF) knockout Hoxb8 cell lines in vitro with the lentivirus (lenti)CRISPR-Cas 9 technique. After their differentiation into neutrophils, the study of their maturation profile, morphology, and effector functions, including NETosis, phagocytosis, and ROS production, is described.
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Acknowledgments
We thank Profs Hans Haecker (University of Utah, USA) and Barbara Walzog (Ludwig Maximilian University of Munich, Germany) for sharing HoxB8 cells and culture protocols with us. This work was supported by the Research into Inflammatory Arthritis Centre Versus Arthritis UK, based in the Universities of Oxford, Glasgow, Birmingham, and Newcastle (22072) and the Kennedy Trust Prize studentship (PhD studentships to J.S.); Oxford-BMS program (fellowship to L.W.); the Chinese Science Council (PhD studentship to Z.A.); and the Wellcome Trust (Investigator Award 209422/Z/17/Z to I.A.U., EvG).
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Salafranca, J., Ai, Z., Wang, L., Udalova, I.A., van Grinsven, E. (2023). Analysis of Neutrophil Morphology and Function Under Genetic Perturbation of Transcription Factors In Vitro. In: Song, Q., Tao, Z. (eds) Transcription Factor Regulatory Networks. Methods in Molecular Biology, vol 2594. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2815-7_6
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DOI: https://doi.org/10.1007/978-1-0716-2815-7_6
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