Use of human placental alkaline phosphatase transgenes to detect somatic mutation in mice in situ.
Use of human placental alkaline phosphatase transgenes to detect somatic mutation in mice in situ.
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DOI：
10.1006/meth.1998.0644
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发表时间：
1998-09
期刊：
影响因子：
4.8
通讯作者：
S. Deprimo;J. Cao;M. Hersh;J. Stringer
中科院分区：
文献类型：
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作者：
S. Deprimo;J. Cao;M. Hersh;J. Stringer
用于原位检测遭受特定突变的细胞的方法对于理解体细胞遗传嵌合现象是有价值的，体细胞遗传嵌合现象是包括癌症在内的多种疾病的基础。这种方法在研究基因表达的变化方面也很有价值，无论是由细胞编程还是由外源性力量引起的，例如暴露于基因毒素或病毒感染。为了改进在动物组织细胞水平上检测遗传变化的方法，我们使用了人胎盘碱性磷酸酶（PLAP）基因。修饰PLAP基因序列，使其不再产生功能性PLAP酶。将突变的PLAP基因置于小鼠基因组中，并研究携带这些突变的PLAP基因的细胞群体以确定将获得PLAP活性的细胞的分数。在培养的细胞和转基因小鼠的组织中研究了突变PLAP基因的自发和诱导逆转。从这些研究中获得的数据显示了原位报告基因如PLAP用于检测组织内变异细胞的效用。
Methods for in situ detection of cells that have suffered a specific mutation would be valuable for understanding somatic genetic mosaicism, a phenomenon that underlies a variety of diseases including cancer. Such methods would also be valuable in studying changes in gene expression, whether programmed by the cells or caused by exogenous forces, such as exposure to genotoxins or infection by a virus. To improve methods for detection of genetic change at the cellular level in animal tissues, we used the human placental alkaline phosphatase (PLAP) gene. The PLAP gene sequence was modified such that it could no longer produce functional PLAP enzyme. Mutant PLAP genes were placed in the mouse genome, and populations of cells carrying these mutant PLAP genes were studied to determine the fraction of cells that would acquire PLAP activity. Spontaneous and induced reversion of mutant PLAP genes was studied in cultured cells and in the tissues of transgenic mice. The data obtained from these studies show the utility of in situ reporter genes such as PLAP for detection of variant cells within a tissue.