Because of shared physiological, anatomical and metabolical features with humans, mice have served for a long time as mammalian disease models. In particular, these last ten years have been the golden age for this favoured model animal. Human and mouse genome projects show that there is 95% genome homology. Spurred by this fact, research attention has shifted from reading these sequences to deciphering the functions of these genes. The 1980s saw the remarkable achievement of homologous recombination in mammalian cell culture systems. Later in the 1990s, innovative gene trapping strategies were developed to enabled random mutagenesis. Today, the goal is to generate more versatile tools to avoid limitations posed by these earlier mutagenesis strategies. Many public and private research centers have united with the aim of mutating all mouse genes. In order to achieve this mutagenesis, the first requirement is a set of practical and efficient viral or plasmid based vectors that can be used globally in the genome. This will be aided by advances in understanding of biological events such as gene transcription, recombination, and embryonic stem cell cycle. In addition, technical improvements such as vector development, precise cell culture assay, and recombinant DNA delivery will also be important. The vector design work in this PhD thesis encompasses 0.00001 % ofthese efforts but may to out to be highly relevant...

Because of shared physiological, anatomical and metabolical features with humans, mice have served for a long time as mammalian disease models. In particular, these last ten years have been the golden age for this favoured model animal. Human and mouse genome projects show that there is 95% genome homology. Spurred by this fact, research attention has shifted from reading these sequences to deciphering the functions of these genes. The 1980s saw the remarkable achievement of homologous recombination in mammalian cell culture systems. Later in the 1990s, innovative gene trapping strategies were developed to enabled random mutagenesis. Today, the goal is to generate more versatile tools to avoid limitations posed by these earlier mutagenesis strategies. Many public and private research centers have united with the aim of mutating all mouse genes. In order to achieve this mutagenesis, the first requirement is a set of practical and efficient viral or plasmid based vectors that can be used globally in the genome. This will be aided by advances in understanding of biological events such as gene transcription, recombination, and embryonic stem cell cycle. In addition, technical improvements such as vector development, precise cell culture assay, and recombinant DNA delivery will also be important. The vector design work in this PhD thesis encompasses 0.00001 % ofthese efforts but may to out to be highly relevant...