Quantification of genomic mutations in murine hematopoietic cells.
Quantification of genomic mutations in murine hematopoietic cells.
复制标题
小鼠造血细胞基因组突变的定量。
DOI：
10.1007/978-1-59745-409-4_28
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Jan,Vijg
中科院分区：
文献类型：
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作者：
Geiger,Hartmut;David,Schleimer;Nattamai,KalpanaJ;Jan,Vijg
维持基因组的稳定性对细胞存活和正常细胞生长至关重要。造血细胞的遗传修饰可能会增加DNA突变积累的固有风险。它经常需要在超生理的氧气水平下培养细胞，这可能导致氧化损伤的增加，以及在超生理的细胞因子浓度下，这可能干扰dna损伤检查点的激活，从而可能导致突变负荷的增加。我们在此描述了一种基于lacz质粒（pUR288）转基因小鼠（小蓝鼠）突变指示菌株的骨髓细胞转导后或在体外或体内选择时监测DNA突变频率的方案。
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.