With the completion of the mouse and human genome sequences, the functional characterization of every mammalian gene and the deciphering of their molecularinteraction network has become the next major challenge, and the mouse genomeprovides a convenient and vital model for these studies. In Gene Knockout Protocols, Second Edition, distinguished contributors with extensive experience in the gene targeting and mouse genetics fields reveal a comprehensive collection of step-by-step laboratory protocols. Emphasizing the many new mutagenesis techniques developed over the last seven years, the volume covers topics such as gene modification in ES cells, stem cell manipulation, the generation of genetically engineered mice, and mutant phenotype analysis. Comprehensive guide to all techniques required for the generation of classical and conditional knockout mouse strains Includes techniques for ES cell establishment, ES cell in vitro differentatio, and nuclear cloning Provides a systematic overview of techniques for mutant phenotyping Covers all gene knockout techniques, including gene targeting, chemical, gene trap, and transposon mutagenesis INDICE: From the contents Overview on Mouse Mutagenesis.- Construction of Gene Targeting Vectors by Recombineering.- Gene Trap Vectors and Mutagenesis.-Chromosome Engineering in ES Cells.- Gene Modification in Embryonic Stem Cells by Single-stranded DNA Oligonucleotides.- Generation of shRNA Transgenic Mice.- Mutagenesis of Mouse Embryonic Stem Cells with Ethylmethanesulfonate.- Gene Targeting in Mouse Embryonic Stem Cells.- Manipulating Mouse Embryonic Stem Cells.- ES Cell Line Establishment.- Generation of Double Knockout Embryonic Stem Cells.- Differentiation Analysis of Pluripotent Mouse Embryonic Stem (ES) Cells in vitro.- Cloning of ES cells and Mice by Nuclear Transfer.- Isolation,Microinjection and Transfer of Mouse Blastocysts.- Aggregation Chimeras: Combining ES Cells, Diploid and Tetraploid Embryos.- VelociMouse: Fully ES cell-derived F0 Generation Mice Obtained from the injection of ES cells into 8-cell Stage Embryos.
- ISBN: 978-1-934115-26-8
- Editorial: Humana
- Encuadernacion: Cartoné
- Páginas: 465
- Fecha Publicación: 01/03/2009
- Nº Volúmenes: 1
- Idioma: Inglés