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Gene knockout protocols / edited by Ralf Kuhn, Wolfgang Wurst. — 2nd ed. — [Totowa, N.J.] : Humana, c2009. – (58.17/M592/v.530)

Contents

    Contents
    
    Preface
    Contributors
    Color Plates
    1. Overview on Mouse Mutagenesis
    PART I: GENE MODIFICATION IN ES CELLS
    2. Construction of Gene-Targeting Vectors by Recombineering
    3. Gene-Trap Vectors and Mutagenesis
    4. Chromosome Engineering in ES Cells 49
    5. Gene Modification in Embryonic Stem Cells by Single-Stranded DNA Oligonucleotides
    6. Generation of shRNA Transgenic Mice
    7. Mutagenesis of Mouse Embryonic Stern Cells with Ethylmethanesulfonate 131
    PART II: STEM CELL MANIPULATION
    8. Gene Targeting in Mouse Embryonic Stem Cells 141
    9. Manipulating Mouse Embryonic Stem Cells 165
    10. ES Cell Line Establishment
    11. Generation of Double-Knockout Embryonic Stem Cells 205
    12. Differentiation Analysis of Pluripotent Mouse Embryonic Stem (ES) Cells In Vitro 219
    13. Cloning of ES Cells and Mice by Nuclear Transfer 251
    PART III: GENETICALLY ENGINEERED Mice
    14. Isolation, Microinjection and Transfer of Mouse Blastocysts
    15. Aggregation Chimeras: Combining ES Cells, Diploid, and Tetraploid Embryos
    16. VelociMouse: Fully ES Cell-Derived F0-Generation Mice Obtained from the Injection of ES Cells into Eight-Cell-Stage Embryos 311
    17. Generation of Cre Recombinase-Expressing Transgenic Mice Using Bacterial Artificial Chromosomes 325
    18. Inducible Cre Mice
    19. Creation and Use of a Cre Recombinase Transgenic Database
    20. Transposon Mutagenesis in Mice
    21. Lentiviral Transgenesis 391
    22. Sperm Cryopreservation and In Vitro Fertilization
    PART IV: PHENOTYPE ANALYSIS23. Influence of Genetic Background on Genetically Engineered Mouse Phenotypes
    24. Pathologic Phenotyping of Mutant Mice
    25. Systemic First-Line Phenotyping 463
    Index