Gene Knockout Protocols

by Tymms, Martin J.; Kola, Ismail

• ISBN13: 9780896035720
• ISBN10: 0896035727
• Format: Hardcover
• Copyright: 2001-01-01
• Publisher: Humana Pr Inc

Summary

Highly skilled investigators with extensive experience in gene targeting and mouse genetics describe their best techniques for the design of targeting constructs and for the analysis of the mouse phenotype. These include embryo transplantation, in vitro embryonic stem cell differentiation, creation of aggregation chimeras, mouse pathology, embryo cryopreservation, and transplantation. State-of-the-art and highly practical, Gene Knockout Protocols not only constitutes an invaluable source of readily reproducible techniques for those just entering the field of gene targeting, but also a key reference for all genetic researchers today.

Table of Contents

Preface	v
Contributors	ix
Overview: Gene Knockouts	1	(10)
Paul J. Hertzog
Ismail Kola
Isolation and Maintenance of Primate Embryonic Stem Cells	11	(8)
Vivienne S. Marshall
Michelle A. Waknitz
James A. Thomson
Gene Targeting in ES Cells	19	(28)
Thomas M. DeChiara
Manipulating Mouse Embryonic Stem Cells	47	(18)
Lino Tessarollo
Gene Targeting in a Centralized Facility	65	(18)
Louise D. Barnett
Frank Kontgen
The LoxP/CRE System and Genome Modification	83	(12)
Trevor J. Wilson
Ismail Kola
Creation and Use of a Cre Recombinase Transgenic Database	95	(12)
Andras Nagy
Lynn Mar
Choice of Mouse Strains for Gene Targeting	107	(14)
Mirelle H. Lahoud
Trevor J. Wilson
Paul J. Hertzog
Isolation, Microinjection, and Transfer of Mouse Blastocysts	121	(14)
Angelika Bonin
Susan W. Reid
Lino Tessarollo
Aggregation Chimeras: Combining ES Cells, Diploid and Tetraploid Embryos	135	(20)
Mika Tanaka
Anna-Katerina Hadjantonakis
Andras Nagy
How to Study Pathologic Phenotypes of Knockout Mice	155	(26)
Roderick T. Bronson
Analysis of Hematopoietic Phenotypes in Knockout Mouse Models	181	(24)
Ivan Bertoncello
Brenda Williams
Isolation of Embryonic Fibroblasts and Their Use in the In Vitro Characterization of Gene Function	205	(12)
Paul J. Hertzog
Influence of Genetic Background on Knockout Mouse Phenotypes	217	(10)
L. Philip Sanford
Suhas Kallapur
Ilona Ormsby
Thomas Doetschman
Lineage Allocation During Early Embryogenesis: Mapping of the Neural Primordia and Application to the Analysis of Mouse Mutants	227	(24)
Gabriel A. Quinlan
Bruce P. Davidson
Patrick P. L. Tam
Generation of Double-Knockout Embryonic Stem Cells	251	(12)
Hein te Riele
Conny Brouwers
Marleen Dekker
In Vitro Differentiation of Embryonic Stem Cells and Analysis of Cellular Phenotypes	263	(24)
Anna M. Wobus
Kaomei Guan
Uta Pich
Embryonic Stem Cells in the Study of Hematopoiesis	287	(14)
Francesca Lazner
Ismail Kola
Elizabeth Stadler
Andrew G. Elefanty
Interferon-Inducible ES Cell Expression Systems	301	(18)
Linda M. Whyatt
Peter D. Rathjen
Transgenic Studies in the Mouse: Improving the Technology Towards a Conditional Temporal and Spatial Approach	319	(16)
Sika Ristevski
In Vivo Libraries of Large Insert Transgenic Mice for Genetic Mapping	335	(16)
Desmond J. Smith
Epigenetic Effects on Transgene Expression	351	(18)
Emma Whitelaw
Heidi Sutherland
Margot Kearns
Hugh Morgan
Linda Weaving
David Garrick
Positional-Candidate Cloning of Genes from Mouse Mutants	369	(12)
Karen B. Avraham
Genetically Engineered Mice: Husbandry and Resources	381	(16)
John J. Sharp
Carol C. Linder
Larry E. Mobraaten
Embryo Cryopreservation for Transgenic Mouse Lines	397	(24)
Jillian M. Shaw
Magosaburo Kasai
Index	421