Aneugen Screening in Optimized Yeast Strains

描述（由申请人提供）：虽然非整倍体是导致 严重的不利人类健康状况，如出生缺陷、精神 发育迟缓、自然流产和癌症，人们对这些疾病知之甚少。 我们接触的化学品和药物可能具有非整倍体性。一期 工作，一种新的酵母菌为基础的染色体不分离测定， 最终用于高通量筛选。这种不分离 检测具有简单的荧光读数，并对已知的非整倍体有反应。在这里， 该测定将适于经济的高通量形式。此外，本发明还提供了一种方法， 提高酵母试验预测值的方法将是 探索了两种方式。第一，降低药物动力学的方法， 酵母和哺乳动物细胞之间的差异，包括1）增加药物 摄取，2）减少药物排泄，和3）模拟哺乳动物的代谢， 将探索测试化合物。二、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-- 将被审查。最后，不分离分析将与 DEL样测定，允许同时评估非整倍体性和 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.