Chromosome Analysis of Single Cells

此应用程序旨在为Brynn Levy博士提供有指导的实验室研究计划 促进他成为一名独立的临床科学家。在他任职前和任职后- 博士学位，利维博士建立了比较基因组杂交(CGH)，一种基于DNA的 染色体异常识别技术是常规临床诊断有价值的工具 临床细胞遗传学以及癌症细胞遗传学。目前的研究目的是使用该工具来 单细胞总非整倍体的检测。因为培养和执行常规是不可行的 对单细胞的细胞遗传学分析，CGH是一种有吸引力的鉴定技术 人类单个细胞中的染色体不平衡。这种方法的应用将为细胞遗传学提供 孕期孕妇外周血中胎儿细胞的产前诊断及临床意义 从2-3天大的胚胎中获取单细胞，用于植入前诊断。长期目标是 因此，(A)提供一种非侵入性的产前诊断方法和(B)提高种植率 通过只移植非整倍体胚胎，可以减少试管受精的风险，并降低随后的流产率。要实现 这些目标，具体目标是：[1]建立常规和DNA微阵列CGH方案 高灵敏度、高特异性、高重复性的单细胞非整倍体分析 用显微操作仪从细胞培养物中提取。[2]确定单电池计算全息的效用 孕妇外周血中胎儿细胞二倍体和非整倍体的预测方法 怀孕了。[3]评估单细胞CGH检测胚胎非整倍体的能力 由于荧光原位杂交(FISH)诊断为非整倍体而被丢弃。[4]评估 嵌合体的真实频率：(A)因FISH诊断为非整倍体而被丢弃的胚胎和(B) 由于遗传性疾病的分子诊断，胚胎被丢弃。利维博士的发展将 以受保护的研究时间、专用的实验室空间和部门来支持他的努力 和机构核心设施。他将在负责任的研究工作中得到指导，他的 通过认真的参与和承诺，将促进发展成为独立的研究人员 他的导师以及西奈山一流的研究和智力环境。

This application is designed to provide Dr. Brynn Levy a program of mentored laboratory research to facilitate his development as an independent clinical scientist. During the course of his pre- and post- doctoral fellowships, Dr. Levy established comparative genomic hybridization (CGH), a DNA-based technique for identifying chromosome abnormalities, to be a valuable clinical diagnostic tool in routine clinical cytogenetics as well as in cancer cytogenetics. Current research is aimed at using this tool for detection of total aneuploidy in single cells. Since it is not feasible to culture and perform conventional cytogenetic analysis on single cells, CGH presents itself as an attractive technique for identifying chromosomal imbalances in single human cells. The application of this method would provide cytogenetic analysis of fetal cells derived from maternal circulation during pregnancy for prenatal diagnosis as well as of single cells obtained from 2-3 day old embryos for preimplantation diagnosis. The long term goals are therefore to (a) provide a non-invasive method of prenatal diagnosis and (b) increase the implantation rate of IVF and decrease the ensuing miscarriage rate by transferring only non-aneuploid embryos. To achieve these goals, the specific aims are: [1] Establish both a regular and DNA microarray CGH protocol which is highly sensitive, specific and reproducible for aneuploidy analysis of single human cells using single cells extracted from cell cultures by micromanipulation apparatus. [2] Determine the utility of the single cell CGH protocols for predicting diploidy and aneuploidy in fetal cells derived from the maternal circulation during pregnancy. [3] Assess the single cell CGH protocols for their ability to detect aneuploidy in embryos discarded because of a Fluorescence in situ hybridization (FISH) diagnosis of aneuploidy. [4] Assess the true frequency of mosaicism in: (A) embryos discarded because of a FISH diagnosis of aneuploidy and (B) embryos discarded because of a molecular diagnosis of an inherited disorder. Dr. Levy's development will be supported in his endeavors with protected research time, dedicated laboratory space, and Departmental and Institutional core facilities. He will be guided in the responsible conduct of research, and his development into an independent researcher will be enhanced by the serious involvement and commitment of his mentor and by the superb research and intellectual environment at Mount Sinai.