Gene Targeting : A Practical Approach

List of contributors Abbreviations 1. Gene targeting vectors for mammalian cells, Paul Hasty and Allan Bradley 1. Introduction 2. Replacement vectors 3. Insertion vectors 4. Maximizing the targeting frequency and selection of targeted clones 5. Selection markers 6. Generating subtle mutations with gene targeting techniques 7. Summary 2. Production of targeted embryonic stem cell clones, W. Wurst and Alexandra L. Joyner 1. Introduction 2. Propagation and maintenance of embryonic stem cells 3. Electroporation of ES cells 4. Screening single colonies for homologous targeting events 3. Analysis of gene transfer in bone marrow stem cells, Keri A. Moore and John W. Belmont 1. Introduction 2. Methods for the transduction of haematopoietic stem cells 3. Tissue culture models of murine and human haematopoietic precursor cell maintenance, growth, and differentiation 4. In vivo models for murine haematopoiesis 5. Human haematopoietic cell analysis 4. Production of chimeras and genetically defined offspring from targeted ES cells, Virginia Papaioannou and Randall Johnson 1. Introduction 2. The starting material 3. Introduction of ES cells into embryos 4. Embryo transfer 5. Chimerism 6. Maintaining a targeted mutation 5. Production of completely ES cell-derived fetuses, Andras Nagy and Janey Rossant 1. Introduction 2. Embryonic stem cells 3. Production of embryo partners 4. Aggregating ES cells with cleavage stage embryos 5. Characterization of mosaicism in diploid or tetraploid chimeras 6. Pre-natal development and perinatal mortality of ES cell diploid and ES cell tetraploid embryos 7. Factors affecting pluripotentiality of ES cells 8. Applications for completely ES cell-derived embryos and fetuses 9. Prospects and limitations 6. Gene and enhancer trap screens in ES cell chimeras, A. Gossler and J. Zachgo 1. Introduction 2. Vector design. 3. Establishment of cell lines carrying reporter gene integrations 4. Production and analysis of chimeras 5. Cloning endogenous genes References Appendix. Suppliers of specialist items IndexJoyner, Alexandra L. is the author of 'Gene Targeting : A Practical Approach', published 1993 under ISBN 9780199634071 and ISBN 0199634076.