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

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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.