Transgenesis Techniques

by Cartwright, Elizabeth J.

• ISBN13: 9781603270182
• ISBN10: 1603270183
• Edition: 3rd
• Format: Hardcover
• Copyright: 2009-06-05
• Publisher: Humana Pr Inc

Summary

One of the major challenges currently facing the scientific community is to understand the function of the multitude of protein-coding genes that were revealed when the human genome was fully sequenced. In Transgenesis Techniques: Principles and Protocols, Third Edition, experts in the field fully update and expand upon the previous edition in order to detail the transgenic techniques currently used to modify the genome. The volume explores classic procedures to genetically modify mice and other model organisms, as well as cutting-edge practices involving microinjection, site-specific recombination systems, cryopreservation, and many other topics. Written in the highly successful Methods in Molecular Biology'„¢ series format, the chapters include brief introductions to their subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes sections, highlighting tips on troubleshooting and avoiding known pitfalls.Comprehensive and state-of-the-art, Transgenesis Techniques: Principles and Protocols, Third Edition is a comprehensive practical guide to the generation of transgenic animals and an invaluable source of information in any lab currently involved in transgenic techniques.

Table of Contents

Preface	p. vii
Contributors	p. xi
Transgenesis in Various Model Systems
Transgenesis in Drosophila melanogaster	p. 3
Transgenesis in Caenorhabditis elegans	p. 21
Transgenesis in Zebrafish with the Tol2 Transposon System	p. 41
Generation of Transgenic Frogs	p. 65
Pronuclear DNA Injection for the Production of Transgenic Rats	p. 73
Transgenesis in the Mouse
Cell-Type-Specific Transgenesis in the Mouse	p. 91
Transgene Design and Delivery into the Mouse Genome: Keys to Success	p. 105
Overexpression Transgenesis in Mouse: Pronuclear Injection	p. 111
Gene-Targeting Vectors	p. 127
Gene Trap: Knockout on the Fast Lane	p. 145
Culture of Murine Embryonic Stem Cells	p. 161
Targeting Embryonic Stem Cells	p. 185
Generation of Chimeras by Microinjection	p. 199
Generation of Chimeras by Morula Aggregation	p. 219
Surgical Techniques for the Generation of Mutant Mice	p. 231
Site-Specific Recombinases for Manipulation of the Mouse Genome	p. 245
Cre Transgenic Mouse Lines	p. 265
Large-Scale Mouse Mutagenesis	p. 275
Dedicated Mouse Production and Husbandry	p. 285
Biological Methods for Archiving and Maintaining Mutant Laboratory Mice. Part I: Conserving Mutant Strains	p. 301
Biological Methods for Archiving and Maintaining Mutant Laboratory Mice. Part II: Recovery and Distribution of Conserved Mutant Strains	p. 321
Index	p. 333
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