In vivo reagents to identify functional noncoding sequences in the TCF7L2 locus

描述（由申请人提供）：最近的一份报告表明，TCF 7L2基因内的遗传变异与3个人群中发生2型糖尿病的风险相关。此后，有几份报告重复并有力地验证了这些初步观察结果，使得寻找因果序列变异和TCF 7L2调节葡萄糖稳态的机制成为糖尿病研究的优先事项。然而，这些报告也表明，致病变异很可能是非蛋白质编码的性质，可能包含在一个92 kb的连锁不平衡块，其中包括TCF 7L2的内含子3和4，使致病序列变异的鉴定极其困难。在这个应用程序中，我们建议测试的假设，TCF7L2的内含子3和4港口进化保守的顺式调控元件负责该基因的组织特异性表达模式。我们将使用生物信息学工具和体内小鼠转基因报告分析技术相结合，以表征内含子3和4内进化上保守的非编码序列，并确定TCF7L2的顺式调节元件，可能含有功能性非编码变异，从而导致糖尿病的风险。这些研究将确定可能隐藏与人类糖尿病相关的致病变异的功能性非编码序列，并将产生关键的分子和体内试剂，可用于设计和测试将TCF 7L2生物学与糖尿病风险联系起来的假设。一些研究表明，TCF 7L2基因的DNA序列变异可能是导致2型糖尿病风险的重要因素。然而，DNA序列的变化可能是在控制该基因何时何地被激活的调控元件中。我们提出了一个合理的策略，以确定这些监管要素。

DESCRIPTION (provided by applicant): A recent report suggested that genetic variations within the TCF7L2 gene are associated with risk to develop type 2 diabetes in 3 populations. Several reports have since replicated and strongly validated these initial observations, making the search for the causal sequence variations and the mechanisms whereby TCF7L2 modulate glucose homeostasis a priority in diabetes research. Nevertheless, these reports also indicate that the causal variation is likely to be non-protein coding in nature, likely contained within a linkage disequilibrium block of 92 kb that includes introns 3 and 4 of TCF7L2, making the identification of the causative sequence variants extremely difficult. In this application, we propose to test the hypothesis that introns 3 and 4 of TCF7L2 harbor evolutionarily conserved cis-regulatory elements responsible for the tissue-specific expression patterns of this gene. We will use a combination of bioinformatic tools and in vivo mouse transgenic report assay technologies to characterize the evolutionarily conserved noncoding sequences within introns 3 and 4, and identify TCF7L2 the cis-regulatory elements that may harbor functional noncoding variation conferring risk to diabetes. These studies will identify the functional noncoding sequences that may harbor the causative variations associated with diabetes in humans and will generate critical molecular and in vivo reagents that can be used to design and test hypotheses that connect TCF7L2 biology to the risk of developing diabetes. Several studies indicate that DNA sequence variation in the TCF7L2 gene may be an important factor conferring risk to develop type 2 diabetes. Nevertheless, the DNA sequence changes are probably in regulatory elements that control where and when this gene is activated. We propose a rational strategy to identify these regulatory elements.