Molecular Detection of DNA Hydroxymethylation for Cancer Screening

 描述(由申请人提供)：该项目将开发一种检测基因组DNA中羟甲基化(HMC)水平的策略。我们将首先将一种选择性地将生物素附着到HMC的方法与我们展示的检测DNA中单一生物素修饰的技术相结合，以演示我们方法的有效性并优化实验条件。然后，我们将把这项技术应用于临床组织样本。首先，我们将测量正常乳腺组织和乳腺癌组织的HMC含量，并将我们的结果与传统方法的结果进行比较。最后，我们将利用我们的方法选择性地分离修饰的DNA的能力，并应用该系统来鉴定LZTS1基因片段的HMC含量的变化，LZTS1基因是一个重要的肿瘤抑制基因。

 DESCRIPTION (provided by applicant): This project will develop a strategy to detect levels of hydroxymethylation (hmC) in genomic DNA. We will first combine a method to selectively attach a biotin to hmC with our demonstrated technique of detecting single biotin modifications in DNA to demonstrate the efficacy of our approach and optimize experimental conditions. We will then apply the technique to clinical tissue samples. First, we will measure the global hmC content of normal breast tissue and of breast carcinoma and compare our findings with results from conventional approaches. Finally, we will utilize the capability of our approach to isolate modified DNA selectively and apply the system to identifying alterations in the hmC content of a fragment of the LZTS1 gene, an important tumor suppressor.