Summary
Maintaining the stability of the genome is critical to cell survival and normal cell growth. Genetic modification of hematopoietic cells might bear an inherent increased risk for the accumulation of DNA mutations. It frequently requires cultivation of the cells under super-physiological oxygen levels, which can result in increased oxidative damage, as well as under super-physiological concentrations of cytokines, which might interfere with DNA-damage checkpoint activation and by this means might result in an increased mutational load. We describe here a protocol for monitoring the frequency of DNA mutations in bone marrow cells post transduction or upon selection either in vitro or in vivo based on the lacZ-plasmid (pUR288) transgenic mouse (small blue mouse) mutation indicator strain.
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Acknowledgments
The author would like to thank Martijn Dollé and Rita Busuttil for their support in learning how to perform the assay and Sina Albert for support in establishing the assay in his laboratory. This work was supported in part by NIH grant R01 HL076604 as well as a New Scholar in Aging grant from The Ellison Medical Foundation.
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Geiger, H., David, S., Nattamai, K.J., Jan, V. (2009). Quantification of Genomic Mutations in Murine Hematopoietic Cells. In: Baum, C. (eds) Genetic Modification of Hematopoietic Stem Cells. Methods In Molecular Biology™, vol 506. Humana Press. https://doi.org/10.1007/978-1-59745-409-4_28
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DOI: https://doi.org/10.1007/978-1-59745-409-4_28
Publisher Name: Humana Press
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