Overview
- Editors:
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Wolfgang Wurst
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Ralf Kühn
- 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
- Written by hands-on experts who have optimized each method for experimental success
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Table of contents (25 protocols)
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Gene Modification in ES Cells
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- Ralf Kühn, Wolfgang Wurst
Pages 1-12
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- Song-Choon Lee, Wei Wang, Pentao Liu
Pages 15-27
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- Silke De-Zolt, Joachim Altschmied, Patricia Ruiz, Harald von Melchner, Frank Schnütgen
Pages 29-47
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- Louise van der Weyden, Charles Shaw-Smith, Allan Bradley
Pages 49-77
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- Marieke Aarts, Marleen Dekker, Rob Dekker, Sandra de Vries, Anja van der Wal, Eva Wielders et al.
Pages 79-99
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- Christiane Hitz, Patricia Steuber-Buchberger, Sabit Delic, Wolfgang Wurst, Ralf Kühn
Pages 101-129
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- Robert Munroe, John Schimenti
Pages 131-138
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Stem Cell Manipulation
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Front Matter
Pages 139-139
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- Lino Tessarollo, Mary Ellen Palko, Keiko Akagi, Vincenzo Coppola
Pages 141-164
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- Eileen Southon, Lino Tessarollo
Pages 165-185
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- Heidrun Kern, Branko Zevnik
Pages 187-204
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- Eva Wielders, Marleen Dekker, Hein te Riele
Pages 205-218
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- Insa S. Schroeder, Cornelia Wiese, Thuy T. Truong, Alexandra Rolletschek, Anna M. Wobus
Pages 219-250
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- Sayaka Wakayama, Satoshi Kishigami, Teruhiko Wakayama
Pages 251-265
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Genetically Engineered Mice
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Front Matter
Pages 266-266
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- Susan W. Reid, Lino Tessarollo
Pages 269-285
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- Mika Tanaka, Anna-Katerina Hadjantonakis, Kristina Vintersten, Andras Nagy
Pages 287-309
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- Thomas M. DeChiara, William T. Poueymirou, Wojtek Auerbach, David Frendewey, George D. Yancopoulos, David M. Valenzuela
Pages 311-324
About this book
Following the completion of the mouse and human genome sequences, a major challengeisthefunctionalcharacterizationofeverymammaliangeneandthedeciph- ing of their molecular interaction network. The mouse offers many advantages for the use of genetics to study human biology and disease, unmatched among other m- mals. Its development, body plan, physiology, behavior, and diseases have much in common, based on the fact that 99% of the human genes have a mouse ortholog. The investigation of gene function using mouse models is based on many years of tech- logical development. In the two decades since gene targeting in murine embryonic stem (ES) cells was first described by Mario Capecchi and colleagues, more than 3000 predesigned mouse mutants have been developed. To date, a variety of mouse mutagenesis techniques, either gene- or phenotype-driven, are used as systematic approaches. The availability of the genome sequence supports gene-driven approaches such as gene-trap and targeted mutagenesis in ES cells, allowing efficient and precise gene disruption. In combination with the use of site-specific DNA recombinases, in particular the Cre/loxP system, gene disruptioncan be directed to specific cell types in conditionalmousemutants. Furthermore,chemicalandtransposonmutagenesisofthe mouse genome enables us to perform phenotype-driven screens for the unbiased identification of phenotype–genotype correlations involved in models of human d- ease. Over the next several years, the mouse genome will be systematically altered, and the techniques for achieving predesigned manipulations will be constantly developed further and improved. The second edition of Gene Knockout Protocols brings together distinguished c-tributorswithextensiveexperienceinthegenetargetingandmousegeneticsfields.
Reviews
From the reviews of the second edition:
"This comprehensive collection of step-by-step protocols in the field of gene targeting and mouse genetics covers such topics as gene modification in embryonic stem cells … and mutant phenotype analysis. … The book will serve as a valuable resource for researchers, scientists, and students interested in these areas. … This is a well-illustrated book … . All researchers interested in gene targeting and gene knockout protocols will find this extremely useful." (Omer Iqbal, Doody’s Review Service, September, 2009)