Analysis of Imprinting Control of the Beckwith Wiedeman Domain

描述（由申请人提供）：本申请要求支持诺拉·恩格尔博士，她是一名初级研究员，以独立研究员的身份开始她的实验室。表观遗传机制的研究将使我们了解基因组如何作为发育蓝图发挥作用，以及基因表达模式的扰动如何导致癌症。本研究的目的是阐明一组印迹基因中导致等位基因特异性基因沉默的表观遗传机制。印迹基因具有亲本特异性的单等位基因表达。本研究的重点是Kcnq1结构域，在该结构域中，一种非编码的反义RNA从父系染色体上表达，并使其邻近基因沉默。我们将首先利用转基因RNA干扰技术来测试kcnq10t的转录是否需要维持该位点的印记。kcnq10t是由Kcnq1的外显子11产生的一种反义非编码RNA。其次，我们将通过染色体构象捕获技术研究Kcnq1结构域调控元件之间的物理相互作用，验证存在等位基因特异性相互作用的假设。在该区域有目标突变的小鼠将与野生型小鼠进行比较。第三个目标是结合生物信息学、生化和体内方法来识别Kcnq1结构域的调控元件。这一建议将为研究开辟道路，提高我们对癌症中印记失调的理解。它也将服务于申请人建立自己作为一个新兴领域的独立调查员的直接职业目标。在良好的职业发展环境的帮助下，通过她将发展的专业知识和合作，申请人将追求成为癌症表观遗传学领域的学术领袖的长期目标。

DESCRIPTION (provided by applicant): This application requests support for Dr. Nora Engel, a junior investigator starting up her laboratory as an independent investigator. The study of epigenetic mechanisms will lead to an understanding of how the genome functions as a developmental blueprint and how perturbations of gene expression patterns can lead to cancer. The goal of this proposal is to elucidate epigenetic mechanisms leading to allele-specific gene silencing in a cluster of imprinted genes. Imprinted genes have parental-specific monoallelic expression. This proposal focuses on the Kcnq1 domain, in which a non-coding, antisense RNA is expressed from the paternal chromosome and silences its neighboring genes. We will first exploit transgenic RNA interference technology to test whether transcription of Kcnq1ot, an imprinted antisense non-coding RNA produced from exon 11 of Kcnq1, is required to maintain the imprinting at this locus. Second, we will study the physical interactions between regulatory elements at the Kcnq1 domain through chromosome conformation capture technology, testing the hypothesis that there are allele-specific interactions. Mice with a targeted mutation in the region will be compared to the wild-type mice. A third aim is to apply a combination of bioinformatic, biochemical and in vivo approaches to identify regulatory elements in the Kcnq1 domain. This proposal will open avenues for research that will improve our understanding of imprinting dysregulation in cancer. It will also serve the immediate career goal of the applicant of establishing herself as an independent investigator in an emerging field. With the help of the excellent career development environment available, and through the expertise and collaborations she will develop, the applicant will pursue the long term goal of becoming an academic leader in cancer epigenetics.