Transgenic ES Cell Differentiation Systems to replace Transgenic Mouse Analysis of Tissue Specific Regulatory Elements

了解基因是如何调控的，对正常发育和癌症等疾病的发展都具有重要意义。对于特定的细胞或组织的发育，必须在严格控制的过程中激活适当的一组基因。目前使用基于细胞的分析和转基因小鼠相结合的方法来确定控制整体基因激活的调控元件，以评估它们活跃的细胞类型。以这种方式分析管制要素的一个主要缺点是涉及小鼠所经历的程序和所需小鼠数量的动物福利问题。在这里，我们建议开发试剂和方案，以取代使用基于体外胚胎干细胞(ES)细胞分化的转基因小鼠。ES细胞很容易接受遗传操作，在适当的培养条件下，有可能发展成许多不同的细胞类型。我们计划系统地检测ES细胞分化过程中的调控元件，并验证可以用这种方法研究的细胞类型，以证明ES细胞分化为评估基因调控功能提供了一种可行的选择。通过提供合适的工具和方案，我们的目标是鼓励更广泛的科学界采用基于胚胎干细胞的系统来分析基因调控，从而大大减少在这类研究中使用的动物数量。此外，这项研究将有助于确定在具有潜在治疗益处的各种细胞类型中特定活跃的调节元件，如免疫细胞或产生胰岛素的胰腺细胞。胚胎干细胞被设计成携带这种细胞类型的特定调控元件，将为开发生产这些细胞的新方案提供强大的工具，因此对于利用胚胎干细胞用于再生医学的前景至关重要。

Understanding how genes are regulated has important implications in both normal development and in the progression of diseases such as cancer. For a specific cell or tissue to develop, the appropriate set of genes must be activated in a tightly controlled process. Regulatory elements that control overall gene activation are currently identified using a combination of cell based assays and transgenic mice to assess the cell types in which they are active. A major drawback of analysing regulatory elements in this way are the animal welfare issues concerning both the procedures that the mice undergo as well as the numbers of mice required. Here, we are proposing to develop reagents and protocols that will replace the use of transgenic mice with in vitro based embryonic stem (ES) cell differentiation assays. ES cells readily undergo genetic manipulation and under appropriate culture conditions have the potential to develop into many different cell types. We plan to systematically test regulatory elements during ES cell differentiation and validate the cell types that can be studied in this way in order to demonstrate that ES cell differentiation provides a viable alternative for assessing gene regulatory function. By providing suitable tools and protocols we aim to encourage the wider scientific community to adopt ES cell based systems for analysing gene regulation thereby dramatically reducing the numbers of animals used in this kind of research. In addition, this study will help to identify regulatory elements that are specifically active in various cell types with potential therapeutic benefits such as immune cells or insulin producing pancreatic cells. ES cells engineered to carry such cell-type specific regulatory elements will provide powerful tools for the development of new protocols for producing these cells and will therefore be vital to harness the promise of embryonic stem cells for regenerative medicine.