Research objectives and content
This project aims to elucidate the biological role of the transcription factor Spl. Spl is expressed ubiquitously and it has been implicated in the expression of many different genes (over 1500 publications). Recently, the Spl gene has been inactivated in the mouse (host laboratory). Spl-deficient mice die at day 10 of gestation, showing that Spl is essential for early embryonic development. However, many putative Spl target genes are expressed normally. Additional results suggest that while Spl is dispensable for growth and differentiation of primitive cells, it is required for viability after differentiation. To test this hypothesis, and to study the role of Spl in fully developed tissues, I propose to generate conditionally targeted Spl mice by flanking exons with loxP sites through homologous recombination, after which the Spl gene can be inactivated by Cre recombinase. To obtain tightly controlled and tissue-specific Cre activity, I will use transgenic mice expressing a hormone-dependent version of Cre in different tissues (host laboratory). Thus, I will be able to inactivate the Spl gene in specific tissues at any developmental stage by injection of artificial hormone, in order to study the role of Spl in gene expression and cellular maintenance in these tissues.
Training content (objective, benefit and expected impact)
In order to carry out this project, I will employ methods like cloning, ES (embryonic stem) cell technology, genetic manipulation of mice, biochemical and immunochemical techniques, as well as state-of-the-art techniques like Representational Difference Analysis (RDA).
Links with industly / industrial lelevance (22)
None are foreseen at the moment.