Aneugen Screening in Optimized Yeast Strains

描述(申请人提供)：尽管非整倍体是导致 严重的不利人类健康状况，如出生缺陷、精神疾病 对于发育迟缓、自然流产和癌症，人们知之甚少 我们接触到的化学物质和药物可能具有非整合性。在第一阶段 工作建立了一种新的基于酿酒酵母的染色体不分离分析方法。 为最终用于高通量筛选而开发。这种不析取 化验有一个简单的荧光读数，并对已知的无核素做出反应。这里, 该分析将被调整为经济的高通量形式。此外, 提高酵母菌检测预测价值的方法将是 从两个方面进行了探索。第一，降低药代动力学的方法 酵母细胞与哺乳动物细胞的差异包括：1)增加药物 摄取，2)减少药物排泄，3)模仿哺乳动物的新陈代谢 将探索测试化合物。二是两种不同类型的“首发” 致敏突变--着丝粒突变和有丝分裂检查点突变-- 将会被检查。最后，非分离分析将与一种 类DEL试验允许同时评估非整倍体和 致癌性。

DESCRIPTION (provided by applicant): Although aneuploidy is responsible for significant adverse human health conditions such as birth defects, mental retardation, spontaneous abortions, and cancer, little is known about the possible aneugenicity of the chemicals and drugs we are exposed to. In Phase I work, a novel Saccharomyces cerevisiae-based chomosome nondisjunction assay was developed for eventual use in high throughput screening. This nondisjunction assay has a simple fluorometric readout and responds to known aneugens. Here, the assay will be adapted to an economical high throughput format. In addition, methods for increasing the predictive value of yeast-based tests will be explored in two ways. First, methods for decreasing the pharmacokinetic differences between yeast and mammalian cells including 1) increasing drug uptake, 2) decreasing drug excretion, and 3) mimicking mammalian metabolism of test compounds will be explored. Second, two different types of "first hit" sensitizing mutations--centromere mutations and mitotic checkpoint mutations-- will be examined. Finally, the nondisjunction assay will be combined with a DEL-like assay to allow simultaneous assessment of both aneugenicity and carcinogenicity.