Bemis Jeffrey C, Hall Nikki E, Dertinger Stephen D
Litron Laboratories, Rochester, NY, USA.
Methods Mol Biol. 2013;1044:51-77. doi: 10.1007/978-1-62703-529-3_3.
In addition to chromosomal damage, assessment of gene mutation is an important part of genotoxicity testing employed during preclinical safety testing. The Pig-a gene mutation assay is based on the loss of function of the Pig-a gene, which results in a lack of cell surface expression of specific proteins that are targeted to the surface by GPI anchors. This cell surface phenotype is readily assessed by flow cytometric analysis of red blood cells. This unit describes a procedure for the collection, processing, and analysis of peripheral blood samples using materials supplied in MutaFlow® kits and a common benchtop flow cytometer.
除了染色体损伤外，基因突变评估是临床前安全性测试中遗传毒性测试的重要组成部分。Pig-a基因突变试验基于Pig-a基因功能的丧失，这导致缺乏通过糖基磷脂酰肌醇（GPI）锚定靶向到表面的特定蛋白质的细胞表面表达。这种细胞表面表型可通过红细胞的流式细胞术分析轻松评估。本单元描述了一种使用MutaFlow®试剂盒中提供的材料和普通台式流式细胞仪收集、处理和分析外周血样本的程序。
