Quantification of genomic mutations in murine hematopoietic cells.
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.
Geiger
Hartmut
H
Department of Medicine, University of Cincinnati, Cincinnati, OH, USA.
David
Schleimer
S
Nattamai
Kalpana J
KJ
Jan
Vijg
V
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D018274:Electroporation; D018014:Gene Transfer Techniques; D023281:Genomics; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009154:Mutation; D016133:Polymerase Chain Reaction
D017931:DNA Primers