PAX6 as a model for synthetic hypervariation studies

人类发育过程中基因表达的调节由称为增强子的遗传开关元件控制。增强子通常位于距离其靶基因很远(100至1000kb)的基因组中，而在发育中重要的基因具有数十个这样的元件，控制着不同细胞类型、不同组织和不同时间的基因表达。这代表了一条我们根本不太了解的基本“生活法则”。为了帮助建立一个平台，系统地解决这样一个深刻的基本问题，即我们的基因组如何控制发育，我们将使用系统的合成基因组方法。这使得很大一部分人类基因组(50万碱基对)的多种变异可以在酵母细胞中以极高的精度构建。然后，这些变异将被精确地传递到脊椎动物的基因组中，以测试经过改造的合成变异是否以及如何在发育过程中发挥作用。在我们的研究中，我们选择了眼睛发育所需的关键基因-PAX6。

The regulation of human gene expression during development is controlled by genetic switch elements called enhancers. Enhancers are often located at huge (100 to 1000kb) genomic distance from their target genes and genes important in development have many tens of such elements controlling gene expression in different cell types, different tissues and at different times. This represents a fundamental "Rule of Life" that we do not understand well at all. To help establish a platform to systematically address such a profound basic question about how our genome controls development, we will use a systematic synthetic genomic approach. This allows multiple variations of a large portion of the human genome (five hundred thousand base pairs) to be constructed with great precision in yeast cells. Then the variants will be precisely delivered to the vertebrate genome to test whether and how the engineered synthetic variants work during development. For our study, we have chosen the key gene required for eye development - PAX6.